WO1998030695A2

WO1998030695A2 - Secreted proteins and polynucleotides encoding them

Info

Publication number: WO1998030695A2
Application number: PCT/US1998/000543
Authority: WO
Inventors: Kenneth Jacobs; John M. Mccoy; Edward R. Lavallie; Lisa A. Racie; David Merberg; Maurice Treacy; Vikki Spaulding; Michael J. Agostino
Original assignee: Genetics Institute, Inc.
Priority date: 1997-01-09
Filing date: 1998-01-09
Publication date: 1998-07-16
Also published as: CA2276092A1; EP0996722A2; WO1998030695A3; AU6022198A; JP2002513279A

Abstract

Novel polynucleotides and the proteins encoded thereby are disclosed.

Description

SECRETED PROTEINS AND POLYNUCLEOTIDES ENCODING THEM

This application is a continuation-in-part of Ser. No. 06/XXX,XXX (converted to a provisional application from non-provisional application Ser. No. 08/780,814), filed January 9, 1997, which is incorporated by reference herein.

FIELD OF THE INVENTION

The present invention provides novel polynucleotides and proteins encoded by such polynucleotides, along with therapeutic, diagnostic and research utilities for these polynucleotides and proteins.

BACKGROUND OF THE INVENTION

Technology aimed at the discovery of protein factors (including e.g., cytokines, such as lymphokines, interferons, CSFs and interleukins) has matured rapidly over the past decade. The now routine hybridization cloning and expression cloning techniques clone novel polynucleotides "directly" in the sense that they rely on information directly related to the discovered protein (i.e., partial DNA/amino acid sequence of the protein in the case of hybridization cloning; activity of the protein in the case of expression cloning). More recent "indirect" cloning techniques such as signal sequence cloning, which isolates DNA sequences based on the presence of a now well-recognized secretory leader sequence motif, as well as various PCR-based or low stringency hybridization cloning techniques, have advanced the state of the art by making available large numbers of DNA/amino acid sequences for proteins that are known to have biological activity by virtue of their secreted nature in the case of leader sequence cloning, or by virtue of the cell or tissue source in the case of PCR-based techniques. It is to these proteins and the polynucleotides encoding them that the present invention is directed. SUMMARY OF THE INVENTION In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:l;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:l from nucleotide 79 to nucleotide 612;

(c) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone BH272_3 deposited under accession number ATCC 98289;

(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone BH272_3 deposited under accession number ATCC 98289;

(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone BH272_3 deposited under accession number ATCC 98289;

(f) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone BH272_3 deposited under accession number ATCC 98289;

(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:2; (h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:2 having biological activity;

(i) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above;

(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above ; and

(k) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(h).

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:l from nucleotide 79 to nucleotide 612; the nucleotide sequence of the full-length protein coding sequence of clone BH272_3 deposited under accession number ATCC

98289; or the nucleotide sequence of the mature protein coding sequence of clone BH272_3 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone BH272_3 deposited under accession number ATCC 98289. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:2 from amino acid 13 to amino acid 164.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:l.

In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:2; (b) the amino acid sequence of SEQ ID NO:2 from amino acid 13 to amino acid 164;

(c) fragments of the amino acid sequence of SEQ ID NO:2; and

(d) the amino acid sequence encoded by the cDNA insert of clone BH272_3 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:2 or the amino acid sequence of SEQ ID NO:2 from amino acid 13 to amino acid 164.

In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:3;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:3 from nucleotide 66 to nucleotide 1256;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:3 from nucleotide 1 to nucleotide 332;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone BP202_3 deposited under accession number ATCC 98289;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone BP202_3 deposited under accession number ATCC 98289;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone BP202_3 deposited under accession number ATCC 98289; (g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone BP202_3 deposited under accession number ATCC 98289;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:4; (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:4 having biological activity;

(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;

(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ; and

(1) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i).

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:3 from nucleotide 66 to nucleotide 1256; the nucleotide sequence of SEQ ID NO:3 from nucleotide 1 to nucleotide 332; the nucleotide sequence of the full-length protein coding sequence of clone BP202_3 deposited under accession number ATCC 98289; or the nucleotide sequence of the mature protein coding sequence of clone BP202_3 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone BP202_3 deposited under accession number ATCC 98289. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:4 from amino acid 1 to amino acid 89. Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:3. In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:4;

(b) the amino acid sequence of SEQ ID NO:4 from amino acid 1 to amino acid 89;

(c) fragments of the amino acid sequence of SEQ ID NO:4; and

(d) the amino acid sequence encoded by the cDNA insert of clone BP202_3 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:4 or the amino acid sequence of SEQ ID NO:4 from amino acid 1 to amino acid 89.

In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:5;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:5 from nucleotide 100 to nucleotide 675; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:5 from nucleotide 208 to nucleotide 489;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CH27_1 deposited under accession number ATCC 98289; (e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CH27_1 deposited under accession number ATCC 98289; (f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CH27_1 deposited under accession number ATCC 98289; (g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CH27_1 deposited under accession number ATCC 98289; (h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:6;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:6 having biological activity;

(j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ; and (1) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i).

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:5 from nucleotide 100 to nucleotide 675; the nucleotide sequence of SEQ ID NO:5 from nucleotide 208 to nucleotide 489; the nucleotide sequence of the full-length protein coding sequence of clone CH27_1 deposited under accession number ATCC 98289; or the nucleotide sequence of the mature protein coding sequence of clone CH27_1 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CH27_1 deposited under accession number ATCC 98289. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:6 from amino acid 39 to amino acid 130.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:5.

(a) the amino acid sequence of SEQ ID NO:6; (b) the amino acid sequence of SEQ ID NO:6 from amino acid 39 to amino acid 130;

(c) fragments of the amino acid sequence of SEQ ID NO:6; and

(d) the amino acid sequence encoded by the cDNA insert of clone CH27_1 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:6 or the amino acid sequence of SEQ ID NO:6 from amino acid 39 to amino acid 130.

NO:7;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:7 from nucleotide 168 to nucleotide 1448;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:7 from nucleotide 440 to nucleotide 941;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CI542_2 deposited under accession number ATCC 98289; (e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CI542_2 deposited under accession number ATCC 98289;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CI542_2 deposited under accession number ATCC 98289;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CI542_2 deposited under accession number ATCC 98289;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:8; (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:8 having biological activity;

(j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:7 from nucleotide 168 to nucleotide 1448; the nucleotide sequence of SEQ ID NO:7 from nucleotide 440 to nucleotide 941; the nucleotide sequence of the full-length protein coding sequence of clone CI542_2 deposited under accession number ATCC 98289; or the nucleotide sequence of the mature protein coding sequence of clone CI542_2 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CI542_2 deposited under accession number ATCC 98289. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:8 from amino acid 170 to amino acid 258.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:7.

(a) the amino acid sequence of SEQ ID NO:8; (b) the amino acid sequence of SEQ ID NO:8 from amino acid 170 to amino acid 258;

(c) fragments of the amino acid sequence of SEQ ID NO:8; and

(d) the amino acid sequence encoded by the cDNA insert of clone CI542_2 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:8 or the amino acid sequence of SEQ ID NO:8 from amino acid 170 to amino acid 258.

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:9;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:9 from nucleotide 112 to nucleotide 1212; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:9 from nucleotide 166 to nucleotide 1212;

(d) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:9 from nucleotide 704 to nucleotide 1033;

(e) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CN483_2 deposited under accession number ATCC 98289;

(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CN483_2 deposited under accession number ATCC 98289;

(g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CN483_2 deposited under accession number

ATCC 98289;

(h) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CN483_2 deposited under accession number ATCC 98289;

(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:10;

(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 10 having biological activity;

(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above; (1) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and

(m) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(j). Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID

NO:9 from nucleotide 112 to nucleotide 1212; the nucleotide sequence of SEQ ID NO:9 from nucleotide 166 to nucleotide 1212; the nucleotide sequence of SEQ ID NO:9 from nucleotide 704 to nucleotide 1033; the nucleotide sequence of the full-length protein coding sequence of clone CN483_2 deposited under accession number ATCC 98289; or the nucleotide sequence of the mature protein coding sequence of clone CN483_2 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CN483_2 deposited under accession number ATCC 98289.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:9.

(a) the amino acid sequence of SEQ ID NO: 10; (b) fragments of the amino acid sequence of SEQ ID NO: 10; and

(c) the amino acid sequence encoded by the cDNA insert of clone CN483_2 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:10. In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:ll;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:ll from nucleotide 234 to nucleotide 1466;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 11 from nucleotide 1 to nucleotide 367; (d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CO1224_2 deposited under accession number ATCC 98289;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CO1224_2 deposited under accession number ATCC 98289;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CO1224_2 deposited under accession number ATCC 98289;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CO1224_2 deposited under accession number ATCC 98289;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 12;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:12 having biological activity; (j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO: 11 from nucleotide 234 to nucleotide 1466; the nucleotide sequence of SEQ ID NO: 11 from nucleotide 1 to nucleotide 367; the nucleotide sequence of the full-length protein coding sequence of clone CO1224_2 deposited under accession number ATCC 98289; or the nucleotide sequence of the mature protein coding sequence of clone C01224_2 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CO1224_2 deposited under accession number ATCC 98289. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:12 from amino acid 1 to amino acid

52.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:ll. In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:12; (b) the amino acid sequence of SEQ ID NO: 12 from amino acid 1 to amino acid 52;

(c) fragments of the amino acid sequence of SEQ ID NO: 12; and

(d) the amino acid sequence encoded by the cDNA insert of clone CO1224_2 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO: 12 or the amino acid sequence of SEQ ID NO:12 from amino acid 1 to amino acid 52.

NO:13;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:13 from nucleotide 845 to nucleotide 1102;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 13 from nucleotide 828 to nucleotide 1111;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CW768_1 deposited under accession number ATCC 98289;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CW768_1 deposited under accession number ATCC 98289;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CW768_1 deposited under accession number ATCC 98289;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CW768_1 deposited under accession number ATCC 98289;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:14;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:14 having biological activity; (j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:13 from nucleotide 845 to nucleotide 1102; the nucleotide sequence of SEQ ID NO:13 from nucleotide 828 to nucleotide 1111; the nucleotide sequence of the full-length protein coding sequence of clone CW768_1 deposited under accession number ATCC 98289; or the nucleotide sequence of the mature protein coding sequence of clone CW768_1 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CW768_1 deposited under accession number ATCC 98289. Other embodiments provide the gene corresponding to the cDNA sequence of SEQ

ID NO:13.

In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of: (a) the amino acid sequence of SEQ ID NO: 14;

(b) fragments of the amino acid sequence of SEQ ID NO:14; and

(c) the amino acid sequence encoded by the cDNA insert of clone C W768_l deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:14.

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:15; (b) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:15 from nucleotide 116 to nucleotide 1081;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:15 from nucleotide 509 to nucleotide 860; (d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CZ268_1 deposited under accession number ATCC 98289;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CZ268_1 deposited under accession number ATCC 98289;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CZ268_1 deposited under accession number ATCC 98289;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CZ268_1 deposited under accession number ATCC 98289;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:16;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:16 having biological activity; (j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:15 from nucleotide 116 to nucleotide 1081; the nucleotide sequence of SEQ ID NO:15 from nucleotide 509 to nucleotide 860; the nucleotide sequence of the full-length protein coding sequence of clone CZ268_1 deposited under accession number ATCC 98289; or the nucleotide sequence of the mature protein coding sequence of clone CZ268_1 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CZ268_1 deposited under accession number ATCC 98289. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:16 from amino acid 153 to amino acid

248.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:15. In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO: 16; (b) the amino acid sequence of SEQ ID NO:16 from amino acid 153 to amino acid 248;

(c) fragments of the amino acid sequence of SEQ ID NO: 16; and

(d) the amino acid sequence encoded by the cDNA insert of clone CZ268_1 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO: 16 or the amino acid sequence of SEQ ID NO:16 from amino acid 153 to amino acid 248.

NO:17;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:17 from nucleotide 348 to nucleotide 2663;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:17 from nucleotide 397 to nucleotide 722;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone DH1308_1 deposited under accession number ATCC 98289;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone DH1308_1 deposited under accession number ATCC 98289;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone DH1308_1 deposited under accession number ATCC 98289;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone DH1308_1 deposited under accession number ATCC 98289;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:18;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:18 having biological activity; (j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:17 from nucleotide 348 to nucleotide 2663; the nucleotide sequence of SEQ ID NO:17 from nucleotide 397 to nucleotide 722; the nucleotide sequence of the full-length protein coding sequence of clone DH1308_1 deposited under accession number ATCC 98289; or the nucleotide sequence of the mature protein coding sequence of clone DH1308_1 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone DH1308_1 deposited under accession number ATCC 98289. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:18 from amino acid 19 to amino acid 125.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:17. In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO: 18;

(b) the amino acid sequence of SEQ ID NO:18 from amino acid 19 to amino acid 125;

(c) fragments of the amino acid sequence of SEQ ID NO: 18; and

(d) the amino acid sequence encoded by the cDNA insert of clone DH1308_1 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:18 or the amino acid sequence of SEQ ID NO:18 from amino acid 19 to amino acid 125.

In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:19;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:19 from nucleotide 158 to nucleotide 3268; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:19 from nucleotide 215 to nucleotide 3268;

(d) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 19 from nucleotide 55 to nucleotide 379;

(e) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone DL185_1 deposited under accession number ATCC 98289;

(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone DL185_1 deposited under accession number ATCC 98289;

(g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone DL185_1 deposited under accession number

ATCC 98289;

(h) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone DL185_1 deposited under accession number ATCC 98289;

(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:20;

(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:20 having biological activity;

(m) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(j). Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:19 from nucleotide 158 to nucleotide 3268; the nucleotide sequence of SEQ ID NO:19 from nucleotide 215 to nucleotide 3268; the nucleotide sequence of SEQ ID NO:19 from nucleotide 55 to nucleotide 379; the nucleotide sequence of the full-length protein coding sequence of clone DL185_1 deposited under accession number ATCC 98289; or the nucleotide sequence of the mature protein coding sequence of clone DL185_1 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone DL185_1 deposited under accession number ATCC 98289. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:20 from amino acid 1 to amino acid 74.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:19.

(a) the amino acid sequence of SEQ ID NO:20;

(b) the amino acid sequence of SEQ ID NO:20 from amino acid 1 to amino acid 74; (c) fragments of the amino acid sequence of SEQ ID NO:20; and

(d) the amino acid sequence encoded by the cDNA insert of clone DL185_1 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:20 or the amino acid sequence of SEQ ID NO:20 from amino acid 1 to amino acid 74.

In certain preferred embodiments, the polynucleotide is operably linked to an expression control sequence. The invention also provides a host cell, including bacterial, yeast, insect and mammalian cells, transformed with such polynucleotide compositions. Also provided by the present invention are organisms that have enhanced, reduced, or modified expression of the gene(s) corresponding to the polynucleotide sequences disclosed herein.

Processes are also provided for producing a protein, which comprise:

(a) growing a culture of the host cell transformed with such polynucleotide compositions in a suitable culture medium; and (b) purifying the protein from the culture.

The protein produced according to such methods is also provided by the present invention. Preferred embodiments include those in which the protein produced by such process is a mature form of the protein. Protein compositions of the present invention may further comprise a pharmaceutically acceptable carrier. Compositions comprising an antibody which specifically reacts with such protein are also provided by the present invention.

Methods are also provided for preventing, treating or ameliorating a medical condition which comprises administering to a mammalian subject a therapeutically effective amount of a composition comprising a protein of the present invention and a pharmaceutically acceptable carrier.

BRIEF DESCRIPTION OF THE DRAWINGS Figures 1 A and IB are schematic representations of the pED6 and pNOTs vectors, respectively, used for deposit of clones disclosed herein.

DETAILED DESCRIPTION ISOLATED PROTEINS AND POLYNUCLEOTIDES Nucleotide and amino acid sequences, as presently determined, are reported below for each clone and protein disclosed in the present application. The nucleotide sequence of each clone can readily be determined by sequencing of the deposited clone in accordance with known methods. The predicted amino acid sequence (both full-length and mature) can then be determined from such nucleotide sequence. The amino acid sequence of the protein encoded by a particular clone can also be determined by expression of the clone in a suitable host cell, collecting the protein and determining its sequence. For each disclosed protein applicants have identified what they have determined to be the reading frame best identifiable with sequence information available at the time of filing. As used herein a "secreted" protein is one which, when expressed in a suitable host cell, is transported across or through a membrane, including transport as a result of signal sequences in its amino acid sequence. "Secreted" proteins include without limitation proteins secreted wholly (e.g., soluble proteins) or partially (e.g. , receptors) from the cell in which they are expressed. "Secreted" proteins also include without limitation proteins which are transported across the membrane of the endoplasmic reticulum.

Clone "BH272 3"

A polynucleotide of the present invention has been identified as clone "BH272_3". BH272_3 was isolated from a human adult ovary (PA-1 teratocarcinoma, pooled retinoic- acid-treated, activin-treated, and untreated tissue) cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. BH272_3 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "BH272_3 protein").

The nucleotide sequence of BH272_3 as presently determined is reported in SEQ ID NO:l. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the BH272_3 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:2.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone BH272_3 should be approximately 1200 bp.

The nucleotide sequence disclosed herein for BH272_3 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. BH272_3 demonstrated at least some similarity with sequences identified as AA030161 (mh86cl l.rl Soares mouse placenta 4NbMP13.5 14.5 Mus

' musculus), AA088849 (zl90d02.s 1 Stratagene colon (#937204) Homo sapiens cDNA clone

51 1875 3'), AA099884 (zl90d02.rl Stratagene colon (#937204) Homo sapiens cDNA clone 51 1875 5' similar to TR G609419 G609419 CHROMOSOME Xπ COSMID 8039), AA102282 (zl81g04.rl Stratagene colon (#937204) Homo sapiens cDNA clone 511062 5' similar to TR G609419 G609419 CHROMOSOME Xπ COSMID 8039), AA150255 (zl07e04.rl Soares pregnant uterus NbHPU Homo sapiens cDNA clone 491646 5' similar to TR G609419 G609419 CHROMOSOME XII COSMID 8039), AA150366 (zl07b05.rl Soares pregnant uterus NbHPU Homo sapiens cDNA clone 491601 5' similar to TR G609419 G609419 CHROMOSOME Xπ COSMID 8039), AF004561 (Homo sapiens p21-Arc mRNA, complete CDs), N98300 (yy68c05.rl Homo sapiens cDNA clone 278696 5' similar to PIR:S51388 S51388 hypothetical protein L8039.15 - yeast), R94679 (yq42gl2.rl Homo sapiens cDNA clone 198502 5'), S51388 (hypothetical protein L8039.15 - yeast), T34031 (EST61558 Homo sapiens cDNA 5' end similar to None), T56617 (yb35d01.sl Homo sapiens cDNA clone 73153 3'), W32065 (zb95f09.rl Soares parathyroid tumor NbHPA Homo sapiens cDNA clone 320585 5' similar to PIR:S51388 S51388 hypothetical protein L8039.15 - yeast), and W58592 (zdl9dl2.rl Soares fetal heart NbHH19W Homo sapiens cDNA clone 341111 5' similar to PIR:S51388 S51388 hypothetical protein L8039.15 - yeast). The predicted amino acid sequence disclosed herein for BH272_3 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted BH272_3 protein demonstrated at least some similarity to sequences identified as AF004561 (p21-Arc [Homo sapiens]) and U19103 (L8039.15 gene product [Saccharomyces cerevisiae]). Based upon sequence similarity, BH272_3 proteins and each similar protein or peptide may share at least some activity.

Clone "BP202 3"

A polynucleotide of the present invention has been identified as clone "BP202_3". BP202_3 was isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. BP202_3 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "BP202_3 protein").

The nucleotide sequence of BP202_3 as presently determined is reported in SEQ ID NO:3. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the BP202_3 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:4.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone BP202_3 should be approximately 1800 bp.

The nucleotide sequence disclosed herein for BP202_3 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. BP202_3 demonstrated at least some similarity with sequences identified as AA112336 (zn68al2.rl Stratagene HeLa cell s3 937216 Homo sapiens cDNA clone 563326 5'), AA505145 (aa65f02.sl NCI_CGAP_GCB1 Homo sapiens cDNA clone AGE:825819 3'), and T05406 (Human cell cycle gene MINI). The predicted amino acid sequence disclosed herein for BP202_3 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted BP202_3 protein demonstrated at least some similarity to sequences identified as R77417 (Human cell cycle protein mini). MINI is a human-derived cell cycle gene encoding an anti-mitotic factor, and is involved in the control of the G2/M phase of the cell cycle. Based upon sequence similarity, BP202_3 proteins and each similar protein or peptide may share at least some activity.

Clone "CH27 1"

A polynucleotide of the present invention has been identified as clone "CH27_1". CH27_1 was isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. CH27_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CH27_1 protein").

The nucleotide sequence of CH27_1 as presently determined is reported in SEQ ID NO:5. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the CH27_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:6.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone CH27_1 should be approximately 1010 bp.

The nucleotide sequence disclosed herein for CH27_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CH27_1 demonstrated at least some similarity with sequences identified as AA086876 (mn93cl0.rl Stratagene mouse Tcell 937311 Mus musculus cDNA clone 5516345' similar to SW SPC3_CANFA P13679 MICROSOMAL SIGNAL PEPTIDASE 21 KD SUBUNIT), C21419 (HUMGS0009728, Human Gene Signature, 3'-directed cDNA sequence), J05069 (Canine 21 kDa signal peptidase subunit mRNA, complete eds), and

W54889 (ma31d03.rl Life Tech mouse brain Mus musculus cDNA clone 312293 5' similar to SW SPC3_CANFA P13679 MICROSOMAL SIGNAL PEPTIDASE 21 KD SUBUNIT). The predicted amino acid sequence disclosed herein for CH27_1 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted CH27_1 protein demonstrated at least some similarity to sequences identified as J05069 (signal peptidase 21 kDa subunit [Canis familiaris]). Based upon sequence similarity, CH27_1 proteins and each similar protein or peptide may share at least some activity. The TopPredll computer program predicts a potential transmembrane domain within the CH27_1 protein sequence centered around amino acid 47 of SEQ ID NO:6.

Clone "CI542 2" A polynucleotide of the present invention has been identified as clone "CI542_2".

CI542_2 was isolated from a human adult brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. CI542_2 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CI542_2 protein").

The nucleotide sequence of CI542_2 as presently determined is reported in SEQ ID NO:7. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the CI542_2 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:8.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone CI542_2 should be approximately 1700 bp.

The nucleotide sequence disclosed herein for CI542_2 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CI542_2 demonstrated at least some similarity with sequences identified as AA114071 (zn76b06.sl Stratagene NT2 neuronal precursor 937230 Homo sapiens cDNA clone 5640833'), AA258839 (zs33hl0.rl NCI_CGAP_GCB1 Homo sapiens cDNA clone IMAGE:6870435'), R06973 (yfl2a09.rl Homo sapiens cDNA clone 1266165'), W20825 (mb91f06.rl Soares mouse p3NMF19.5 Mus musculus cDNA clone 336803 5'), W93955 (zd98f03.rl Soares fetal heart NbHH19W Homo sapiens cDNA clone 357533 5'), and W94010 (zd98f03.sl Soares fetal heart NbHH19W Homo sapiens cDNA clone 357533 3')- Based upon sequence similarity, CI542_2 proteins and each similar protein or peptide may share at least some activity.

Clone "CN483 2"

A polynucleotide of the present invention has been identified as clone "CN483_2". CN483_2 was isolated from a human fetal brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. CN483_2 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CN483_2 protein").

The nucleotide sequence of CN483_2 as presently determined is reported in SEQ ID NO:9. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the CN483_2 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:10. Amino acids 6 to 18 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 19, or are a transmembrane domain. The EcoRI/NotI restriction fragment obtainable from the deposit containing clone

CN483_2 should be approximately 2000 bp.

The nucleotide sequence disclosed herein for CN483_2 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CN483_2 demonstrated at least some similarity with sequences identified as AAl 12426 (zm27d08.rl Stratagene pancreas (#937208) Homo sapiens cDNA clone 526863 5'), AA475778 (vh23f01.rl Soares mouse mammary gland NbMMG Mus musculus), H51262 (yp83b07.sl Homo sapiens cDNA clone 194005 3'), N57441 (yw88b09.rl Homo sapiens cDNA clone 259289 5'), W02460 (za47c01.rl Soares fetal liver spleen 1NFLS Homo sapiens cDNA clone 295680 5'), and X92871 (X.laevis mRNA for an unknown transmembrane protein). The predicted amino acid sequence disclosed herein for CN483_2 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted CN483_2 protein demonstrated at least some similarity to sequences identified as X92871 (unknown transmembrane protein [Xenopus laevis]). Based upon sequence similarity, CN483_2 proteins and each similar protein or peptide may share at least some activity. The TopPredll computer program predicts four potential transmembrane domains within the CN483_2 protein sequence centered around amino acids 190, 220, 300, and 350 of SEQ ID NO:10, respectively.

Clone "CQ1224 2"

A polynucleotide of the present invention has been identified as clone "C01224_2". C01224_2 was isolated from a human adult brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. C01224_2 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "C01224_2 protein").

The nucleotide sequence of C01224_2 as presently determined is reported in SEQ ID NO:ll. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the C01224_2 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:12.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone C01224_2 should be approximately 2300 bp. The nucleotide sequence disclosed herein for C01224_2 was searched against the

GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. No hits were found in the database.

Clone "CW768 1" A polynucleotide of the present invention has been identified as clone "CW768_1 ".

CW768_1 was isolated from a human fetal brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. CW768_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CW768_1 protein").

The nucleotide sequence of CW768_1 as presently determined is reported in SEQ ID NO:13. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the CW768_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:14.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone CW768_1 should be approximately 1200 bp.

The nucleotide sequence disclosed herein for CW768_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CW768_1 demonstrated at least some similarity with sequences identified as AA425283 (zw46d02.sl Soares total fetus Nb2HF8 9w Homo sapiens cDNA clone 773091 3' similar to contains Alu repetitive element;contains element MER22 repetitive element), N27362 (yw52g05.sl Homo sapiens cDNA clone 255896 3' similar to contains Alu repetitive element;contains element MER1 repetitive element), Q60793 (Human brain Expressed Sequence Tag EST01598), U52112 (Human Xq28 genomic DNA in the region of the LICAM locus containing the genes for neural cell adhesion molecule LI (LICAM), arginine-vasopressin receptor (AVPR2), Cl pi 15 (Cl)), and Z82201 (Human DNA sequence from PAC 345P10 on chromosome 22ql2-qter contains ESTs and STS and polymorphic CA repeat D22S927). The predicted amino acid sequence disclosed herein for CW768_1 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted CW768_1 protein demonstrated at least some similarity to sequences identified as L24521 (transformation-related protein [Homo sapiens]). Based upon sequence similarity, CW768_1 proteins and each similar protein or peptide may share at least some activity. The nucleotide sequence of CW768_1 indicates that it may contain an Alu repetitive element.

Clone "CZ268 1" A polynucleotide of the present invention has been identified as clone "CZ268_1 ".

CZ268_1 was isolated from a human adult testes cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. CZ268_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CZ268_1 protein").

The nucleotide sequence of CZ268_1 as presently determined is reported in SEQ ID NO:15. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the CZ268_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:16.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone CZ268_1 should be approximately 1500 bp.

The nucleotide sequence disclosed herein for CZ268_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CZ268_1 demonstrated at least some similarity with sequences identified as AA108889 (ml56bll.rl Stratagene mouse testis (#937308) Mus musculus cDNA clone 5159975' similar to SW:MDP1_PIG P22412 MICROSOMAL DIPEPTIDASE PRECURSOR), D13138 (Human mRNA for dipeptidase), H10870 (ym06a06.rl Homo sapiens cDNA clone 47018 5' similar to SP MDP1_PIG P22412 MICROSOMAL DIPEPTIDASE PRECURSOR), J05257 (Homo sapiens (clones MDP4, MDP7) microsomal dipeptidase (MDP) mRNA, complete eds), Q35201 (pDHP2 coding sequence), and R20620 (yf59a05.sl Homo sapiens cDNA clone 26189 3' similar to SP MDP1_RABIT P31429 MICROSOMAL DIPEPTIDASE PRECURSOR). The predicted amino acid sequence disclosed herein for CZ268_1 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted CZ268_1 protein demonstrated at least some similarity to sequences identified as D13137 (dipeptidase precursor [Homo sapiens]), J05257 (microsomal dipeptidase [Homo sapiens]), and R13857 (Microsomal dipeptidase). Microsomal dipeptidase proteins may be associated with brush-border membranes. Based upon sequence similarity, CZ268_1 proteins and each similar protein or peptide may share at least some activity.

The TopPredll computer program predicts a potential transmembrane domain within the CZ268_1 protein sequence centered around amino acid 312 of SEQ ID NO:16. Amino acids 151 to 163 of SED ID NO:16 are a putative leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 164, or are a transmembrane domain.

Clone "DH1308 1"

A polynucleotide of the present invention has been identified as clone "DH1308_1". DH1308_1 was isolated from a human fetal brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. DH1308_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "DH1308_1 protein").

The nucleotide sequence of DH1308_1 as presently determined is reported in SEQ ID NO:17. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the DH1308_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:18. The EcoRI/NotI restriction fragment obtainable from the deposit containing clone

DH1308_1 should be approximately 3300 bp.

The nucleotide sequence disclosed herein for DH1308_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. DH1308_1 demonstrated at least some similarity with sequences identified as W13942 (mb23h02.rl Soares mouse p3NMF19.5 Mus musculus cDNA clone 330291 5'). Based upon sequence similarity, DH1308_1 proteins and each similar protein or peptide may share at least some activity.

Clone "DL185 1"

A polynucleotide of the present invention has been identified as clone "DL185_1". DL185_1 was isolated from a human adult brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. DL185_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "DL185_1 protein").

The nucleotide sequence of DL185_1 as presently determined is reported in SEQ ID NO:19. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the DL185_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:20. Amino acids 7 to 19 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 20, or are a transmembrane domain.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone DL185_1 should be approximately 3480 bp.

The nucleotide sequence disclosed herein for DL185_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. DL185_1 demonstrated at least some similarity with sequences identified as D87248 (Rat mRNA for NB-3, complete eds), U35371 (Rattus norvegicus neural cell adhesion protein BIG-2 precursor (BIG-2) mRNA, complete eds), and X99043

(M.musculus mRNA for brain-derived immunoglobulin superfamily molecule BIG-2). The predicted amino acid sequence disclosed herein for DL185_1 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted DL185_1 protein demonstrated at least some similarity to sequences identified as D87248 (NB-3 [Rattus norvegicus]), L01991 (neuronal glycoprotein

[Mus musculus]), R63759 (Human contactin (EMBL Accession #Z21488)), and X99043 (brain-derived immunoglobulin superfamily molecule [Mus musculus]). Contactin proteins mediate cell surface interactions during nervous system development and are attached to the membrane by a glycosylphosphatidylinositol anchor. Based upon sequence similarity, DL185_1 proteins and each similar protein or peptide may share at least some activity. The TopPredll computer program predicts an additional potential transmembrane domain within the DL185_1 protein sequence centered around amino acid 1010 of SEQ ID NO:20. The DL185_1 protein sequence also contains an ABC transporter motif at amino acid 639 of SEQ ID NO:20. ABC transporters are involved in an extracellular transport pathway for proteins that lack signal sequences.

Deposit of Clones

Clones BH272_3, BP202_3, CH27_1, CI542_2, CN483_2, C01224_2, CW768_1, CZ268_1, DH1308_1 and DL185_1 were deposited on January 8, 1997 with the American Type Culture Collection as an original deposit under the Budapest Treaty and were given the accession number ATCC 98289, from which each clone comprising a particular polynucleotide is obtainable. All restrictions on the availability to the public of the deposited material will be irrevocably removed upon the granting of the patent, except for the requirements specified in 37 C.F.R. § 1.808(b).

Each clone has been transfected into separate bacterial cells (E. coli) in this composite deposit. Each clone can be removed from the vector in which it was deposited by performing an EcoRI/NotI digestion (5' site, EcoRI; 3' site, NotI) to produce the appropriate fragment for such clone. Each clone was deposited in either the pED6 or pNOTs vector depicted in Fig. 1. The pED6dpc2 vector ("pED6") was derived from pEDόdpcl by insertion of a new polylinker to facilitate cDNA cloning (Kaufman et al, 1991, Nucleic Acids Res. 19: 4485-4490); the pNOTs vector was derived from pMT2 (Kaufman et al, 1989, Mol. Cell. Biol. 9: 946-958) by deletion of the DHFR sequences, insertion of a new polylinker, and insertion of the M13 origin of replication in the Clal site. In some instances, the deposited clone can become "flipped" (i.e., in the reverse orientation) in the deposited isolate. In such instances, the cDNA insert can still be isolated by digestion with EcoRI and NotI. However, NotI will then produce the 5' site and EcoRI will produce the 3' site for placement of the cDNA in proper orientation for expression in a suitable vector. The cDNA may also be expressed from the vectors in which they were deposited.

Bacterial cells containing a particular clone can be obtained from the composite deposit as follows:

An oligonucleotide probe or probes should be designed to the sequence that is known for that particular clone. This sequence can be derived from the sequences provided herein, or from a combination of those sequences. The sequence of the oligonucleotide probe that was used to isolate each full-length clone is identified below, and should be most reliable in isolating the clone of interest.

Clone Probe Sequence

BH272_3 SEQ ID NO:21

BP202_3 SEQ ID NO:22

CH27_1 SEQ ID NO:23

CI542_2 SEQ ID NO:24 CN483_2 SEQ ID NO:25

C01224_2 SEQ ID NO:26

CW768_1 SEQ ID NO:27

CZ268_1 SEQ ID NO:28

DH1308_1 SEQ ID NO:29 DL185_1 SEQ ID NO:30

In the sequences listed above which include an N at position 2, that position is occupied in preferred probes/primers by a biotinylated phosphoaramidite residue rather than a nucleotide (such as , for example, that produced by use of biotin phosphoramidite (1- dimethoxytrityloxy-2-(N-biotinyl-4-aminobutyl)-propyl-3-0-(2-cyanoethyl)-(N,N- diisopropyl)-phosphoramadite) (Glen Research, cat. no. 10-1953)).

The design of the oligonucleotide probe should preferably follow these parameters:

(a) It should be designed to an area of the sequence which has the fewest ambiguous bases ("N's"), if any;

(b) It should be designed to have a T_m of approx. 80 ° C (assuming 2° for each A or T and 4 degrees for each G or C).

The oligonucleotide should preferably be labeled with g-³²P ATP (specific activity 6000 Ci/mmole) and T4 polynucleotide kinase using commonly employed techniques for labeling oligonucleotides. Other labeling techniques can also be used. Unincorporated label should preferably be removed by gel filtration chromatography or other established methods. The amount of radioactivity incorporated into the probe should be quantitated by measurement in a scintillation counter. Preferably, specific activity of the resulting probe should be approximately 4e+6 dpm/pmole. The bacterial culture containing the pool of full-length clones should preferably be thawed and 100 μl of the stock used to inoculate a sterile culture flask containing 25 ml of sterile L-broth containing ampicillin at 100 μg/ml. The culture should preferably be grown to saturation at 37°C, and the saturated culture should preferably be diluted in fresh L-broth. Aliquots of these dilutions should preferably be plated to determine the dilution and volume which will yield approximately 5000 distinct and well-separated colonies on solid bacteriological media containing L-broth containing ampicillin at 100 μg/ml and agar at 1.5% in a 150 mm petri dish when grown overnight at 37°C. Other known methods of obtaining distinct, well-separated colonies can also be employed. Standard colony hybridization procedures should then be used to transfer the colonies to nitrocellulose filters and lyse, denature and bake them.

The filter is then preferably incubated at 65°C for 1 hour with gentle agitation in 6X SSC (20X stock is 175.3 g NaCl/liter, 88.2 g Na citrate /liter, adjusted to pH 7.0 with NaOH) containing 0.5% SDS, 100 μg/ml of yeast RNA, and 10 mM EDTA (approximately 10 mL per 150 mm filter). Preferably, the probe is then added to the hybridization mix at a concentration greater than or equal to le+6 dpm/mL. The filter is then preferably incubated at 65°C with gentle agitation overnight. The filter is then preferably washed in 500 mL of 2X SSC/0.5% SDS at room temperature without agitation, preferably followed by 500 mL of 2X SSC/0.1% SDS at room temperature with gentle shaking for 15 minutes. A third wash with 0.1X SSC/0.5% SDS at 65°C for 30 minutes to 1 hour is optional. The filter is then preferably dried and subjected to autoradiography for sufficient time to visualize the positives on the X-ray film. Other known hybridization methods can also be employed.

The positive colonies are picked, grown in culture, and plasmid DNA isolated using standard procedures. The clones can then be verified by restriction analysis, hybridization analysis, or DNA sequencing.

Fragments of the proteins of the present invention which are capable of exhibiting biological activity are also encompassed by the present invention. Fragments of the protein may be in linear form or they may be cyclized using known methods, for example, as described in H.U. Saragovi, et al, Bio /Technology 10, 773-778 (1992) and in R.S.

McDowell, et al, J. Amer. Chem. Soc. 114, 9245-9253 (1992), both of which are incorporated herein by reference. Such fragments may be fused to carrier molecules such as immunoglobulins for many purposes, including increasing the valency of protein binding sites. For example, fragments of the protein may be fused through "linker" sequences to the Fc portion of an immunoglobulin. For a bivalent form of the protein, such a fusion could be to the Fc portion of an IgG molecule. Other immunoglobulin isotypes may also be used to generate such fusions. For example, a protein - IgM fusion would generate a decavalent form of the protein of the invention. The present invention also provides both full-length and mature forms of the disclosed proteins. The full-length form of the such proteins is identified in the sequence listing by translation of the nucleotide sequence of each disclosed clone. The mature form of such protein may be obtained by expression of the disclosed full-length polynucleotide (preferably those deposited with ATCC) in a suitable mammalian cell or other host cell. The sequence of the mature form of the protein may also be determinable from the amino acid sequence of the full-length form.

The present invention also provides genes corresponding to the polynucleotide sequences disclosed herein. "Corresponding genes" are the regions of the genome that are transcribed to produce the mRNAs from which cDNA polynucleotide sequences are derived and may include contiguous regions of the genome necessary for the regulated expression of such genes. Corresponding genes may therefore include but are not limited to coding sequences, 5' and 3' untranslated regions, alternatively spliced exons, introns, promoters, enhancers, and silencer or suppressor elements. The corresponding genes can be isolated in accordance with known methods using the sequence information disclosed herein. Such methods include the preparation of probes or primers from the disclosed sequence information for identification and/or amplification of genes in appropriate genomic libraries or other sources of genomic materials. An "isolated gene" is a gene that has been separated from the adjacent coding sequences, if any, present in the genome of the organism from which the gene was isolated. Organisms that have enhanced, reduced, or modified expression of the gene(s) corresponding to the polynucleotide sequences disclosed herein are provided. The desired change in gene expression can be achieved through the use of antisense polynucleotides or ribozymes that bind and /or cleave the mRNA transcribed from the gene (Albert and Morris, 1994, Trends Pharmacol. Sci. 15(7): 250-254; Lavarosky et al, 1997, Biochem. Mol. Med. 62(1): 11-22; and Hampel, 1998, Prog. Nucleic Acid Res. Mol. Biol. 58: 1-

39; all of which are incorporated by reference herein). Transgenic animals that have multiple copies of the gene(s) corresponding to the polynucleotide sequences disclosed herein, preferably produced by transformation of cells with genetic constructs that are stably maintained within the transformed cells and their progeny, are provided. Transgenic animals that have modified genetic control regions that increase or reduce gene expression levels, or that change temporal or spatial patterns of gene expression, are also provided (see European Patent No. 0 649464 Bl, incorporated by reference herein). In addition, organisms are provided in which the gene(s) corresponding to the polynucleotide sequences disclosed herein have been partially or completely inactivated, through insertion of extraneous sequences into the corresponding gene(s) or through deletion of all or part of the corresponding gene(s). Partial or complete gene inactivation can be accomplished through insertion, preferably followed by imprecise excision, of transposable elements (Plasterk, 1992, Bioessays 14(9): 629-633; Zwaal et al, 1993, Proc. Natl. Acad. Sci. USA 90(16): 7431-7435; Clark et al, 1994, Proc. Natl. Acad. Sci. USA 91(2): 719-722; all of which are incorporated by reference herein), or through homologous recombination, preferably detected by positive/negative genetic selection strategies (Mansour et al, 1988, Nature 336: 348-352; U.S. Patent Nos. 5,464,764; 5,487,992; 5,627,059; 5,631,153; 5,614, 396; 5,616,491; and 5,679,523; all of which are incorporated by reference herein). These organisms with altered gene expression are preferably eukaryotes and more preferably are mammals. Such organisms are useful for the development of non-human models for the study of disorders involving the corresponding gene(s), and for the development of assay systems for the identification of molecules that interact with the protein product(s) of the corresponding gene(s). Where the protein of the present invention is membrane-bound (e.g., is a receptor), the present invention also provides for soluble forms of such protein. In such forms part or all of the intracellular and transmembrane domains of the protein are deleted such that the protein is fully secreted from the cell in which it is expressed. The intracellular and transmembrane domains of proteins of the invention can be identified in accordance with known techniques for determination of such domains from sequence information.

Proteins and protein fragments of the present invention include proteins with amino acid sequence lengths that are at least 25%(more preferably at least 50%, and most preferably at least 75%) of the length of a disclosed protein and have at least 60% sequence identity (more preferably, at least 75% identity; most preferably at least 90% or 95% identity) with that disclosed protein, where sequence identity is determined by comparing the amino acid sequences of the proteins when aligned so as to maximize overlap and identity while minimizing sequence gaps. Also included in the present invention are proteins and protein fragments that contain a segment preferably comprising 8 or more (more preferably 20 or more, most preferably 30 or more) contiguous amino acids that shares at least 75% sequence identity (more preferably, at least 85% identity; most preferably at least 95% identity) with any such segment of any of the disclosed proteins. Species homologs of the disclosed polynucleotides and proteins are also provided by the present invention. As used herein, a "species homologue" is a protein or polynucleotide with a different species of origin from that of a given protein or polynucleotide, but with significant sequence similarity to the given protein or polynucleotide, as determined by those of skill in the art. Species homologs may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source from the desired species. The invention also encompasses allelic variants of the disclosed polynucleotides or proteins; that is, naturally-occurring alternative forms of the isolated polynucleotide which also encode proteins which are identical, homologous, or related to that encoded by the polynucleotides .

The invention also includes polynucleotides with sequences complementary to those of the polynucleotides disclosed herein.

The present invention also includes polynucleotides capable of hybridizing under reduced stringency conditions, more preferably stringent conditions, and most preferably highly stringent conditions, to polynucleotides described herein. Examples of stringency conditions are shown in the table below: highly stringent conditions are those that are at least as stringent as, for example, conditions A-F; stringent conditions are at least as stringent as, for example, conditions G-L; and reduced stringency conditions are at least as stringent as, for example, conditions M-R.

* The hybrid length is that anticipated for the hybridized regιon(s) of the hybridizing polynucleotides When hybridizing a polynucleotide to a target polynucleotide of unknown sequence, the hybrid length is assumed to be that of the hybridizing polynucleotide When polynucleotides of known sequence are hybridized, the hybrid length can be determined by aligning the sequences of the polynucleotides and identifying the region or regions of optimal sequence complementarity

⁺ SSPE (lxSSPE is 0 15M NaCI, lOmM NaH₂P0₄, and 1 25mM EDTA, pH 74) can be substituted for SSC (lxSSC is 0 15M NaCI and 15mM sodium citrate) in the hybridization and wash buffers, washes are performed for 15 minutes after hybridization is complete

*T_B - T_R The hybridization temperature for hybrids anticipated to be less than 50 base pairs in length should be 5-10°C less than the melting temperature (T_m) of the hybrid, where T_m is determined according to the following equations For hybrids less than 18 base pairs m length, T_m(^cC) = 2(# of A + T bases) + 4(# of G + C bases) For hybrids between 18 and 49 base pairs in length, T_m(°C) = 81 5 + 166(log₁₀[Na⁺]) + 041(%G+C) - (600/N), where N is the number of bases in the hybrid, and [Na⁺] is the concentration of sodium ions in the hybridization buffer ([Na⁺] for lxSSC = 0 165 M) Additional examples of stringency conditions for polynucleotide hybridization are provided in Sambrook, J., E.F. Fritsch, and T. Maniatis, 1989, Molecular Cloning: A

Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, chapters 9 and 11, and Current Protocols in Molecular Biology, 1995, F.M. Ausubel et al., eds., John Wiley & Sons, Inc., sections 2.10 and 6.3-6.4, incorporated herein by reference.

Preferably, each such hybridizing polynucleotide has a length that is at least 25%(more preferably at least 50%, and most preferably at least 75%) of the length of the polynucleotide of the present invention to which it hybridizes, and has at least 60% sequence identity (more preferably, at least 75% identity; most preferably at least 90% or 95% identity) with the polynucleotide of the present invention to which it hybridizes, where sequence identity is determined by comparing the sequences of the hybridizing polynucleotides when aligned so as to maximize overlap and identity while minimizing sequence gaps.

The isolated polynucleotide of the invention may be operably linked to an expression control sequence such as the pMT2 or pED expression vectors disclosed in Kaufman et al, Nucleic Acids Res. 19, 4485-4490 (1991), in order to produce the protein recombinantly. Many suitable expression control sequences are known in the art. General methods of expressing recombinant proteins are also known and are exemplified in R. Kaufman, Methods in Enzymology 185, 537-566 (1990). As defined herein "operably linked" means that the isolated polynucleotide of the invention and an expression control sequence are situated within a vector or cell in such a way that the protein is expressed by a host cell which has been transformed (transfected) with the ligated polynucleotide/expression control sequence.

A number of types of cells may act as suitable host cells for expression of the protein. Mammalian host cells include, for example, monkey COS cells, Chinese Hamster

Ovary (CHO) cells, human kidney 293 cells, human epidermal A431 cells, human Colo205 cells, 3T3 cells, CV-1 cells, other transformed primate cell lines, normal diploid cells, cell strains derived from in vitro culture of primary tissue, primary explants, HeLa cells, mouse L cells, BHK, HL-60, U937, HaK or Jurkat cells. Alternatively, it may be possible to produce the protein in lower eukaryotes such as yeast or in prokaryotes such as bacteria. Potentially suitable yeast strains include Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces strains, Candida, or any yeast strain capable of expressing heterologous proteins. Potentially suitable bacterial strains include Escherichia coli, Bacillus subtilis, Salmonella typhimurium, or any bacterial strain capable of expressing heterologous proteins. If the protein is made in yeast or bacteria, it may be necessary to modify the protein produced therein, for example by phosphorylation or glycosylation of the appropriate sites, in order to obtain the functional protein. Such covalent attachments may be accomplished using known chemical or enzymatic methods.

The protein may also be produced by operably linking the isolated polynucleotide of the invention to suitable control sequences in one or more insect expression vectors, and employing an insect expression system. Materials and methods for baculovirus/insect cell expression systems are commercially available in kit form from, e.g., Invitrogen, San Diego, California, U.S.A. (the MaxBac® kit), and such methods are well known in the art, as described in Summers and Smith, Texas Agricultural Experiment Station Bulletin No. 1555 (1987), incorporated herein by reference. As used herein, an insect cell capable of expressing a polynucleotide of the present invention is "transformed." The protein of the invention may be prepared by culturing transformed host cells under culture conditions suitable to express the recombinant protein. The resulting expressed protein may then be purified from such culture (i.e., from culture medium or cell extracts) using known purification processes, such as gel filtration and ion exchange chromatography. The purification of the protein may also include an affinity column containing agents which will bind to the protein; one or more column steps over such affinity resins as concanavalin A-agarose, heparin-toyopearl® or Cibacrom blue 3GA Sepharose®; one or more steps involving hydrophobic interaction chromatography using such resins as phenyl ether, butyl ether, or propyl ether; or immunoaffinity chromatography. Alternatively, the protein of the invention may also be expressed in a form which will facilitate purification. For example, it may be expressed as a fusion protein, such as those of maltose binding protein (MBP), glutathione-S-transferase (GST) or thioredoxin (TRX). Kits for expression and purification of such fusion proteins are commercially available from New England BioLab (Beverly, MA), Pharmacia (Piscataway, NJ) and InVitrogen, respectively. The protein can also be tagged with an epitope and subsequently purified by using a specific antibody directed to such epitope. One such epitope ("Flag") is commercially available from Kodak (New Haven, CT).

Finally, one or more reverse-phase high performance liquid chromatography (RP- HPLC) steps employing hydrophobic RP-HPLC media, e.g., silica gel having pendant methyl or other aliphatic groups, can be employed to further purify the protein. Some or all of the foregoing purification steps, in various combinations, can also be employed to provide a substantially homogeneous isolated recombinant protein. The protein thus purified is substantially free of other mammalian proteins and is defined in accordance with the present invention as an "isolated protein."

The protein of the invention may also be expressed as a product of transgenic animals, e.g., as a component of the milk of transgenic cows, goats, pigs, or sheep which are characterized by somatic or germ cells containing a nucleotide sequence encoding the protein. The protein may also be produced by known conventional chemical synthesis.

Methods for constructing the proteins of the present invention by synthetic means are known to those skilled in the art. The synthetically-constructed protein sequences, by virtue of sharing primary, secondary or tertiary structural and /or conformational characteristics with proteins may possess biological properties in common therewith, including protein activity. Thus, they may be employed as biologically active or immunological substitutes for natural, purified proteins in screening of therapeutic compounds and in immunological processes for the development of antibodies.

The proteins provided herein also include proteins characterized by amino acid sequences similar to those of purified proteins but into which modification are naturally provided or deliberately engineered. For example, modifications in the peptide or DNA sequences can be made by those skilled in the art using known techniques. Modifications of interest in the protein sequences may include the alteration, substitution, replacement, insertion or deletion of a selected amino acid residue in the coding sequence. For example, one or more of the cysteine residues may be deleted or replaced with another amino acid to alter the conformation of the molecule. Techniques for such alteration, substitution, replacement, insertion or deletion are well known to those skilled in the art (see, e.g., U.S. Patent No.4,518,584). Preferably, such alteration, substitution, replacement, insertion or deletion retains the desired activity of the protein.

Other fragments and derivatives of the sequences of proteins which would be expected to retain protein activity in whole or in part and may thus be useful for screening or other immunological methodologies may also be easily made by those skilled in the art given the disclosures herein. Such modifications are believed to be encompassed by the present invention. USES AND BIOLOGICAL ACTIVITY

The polynucleotides and proteins of the present invention are expected to exhibit one or more of the uses or biological activities (including those associated with assays cited herein) identified below. Uses or activities described for proteins of the present invention may be provided by administration or use of such proteins or by administration or use of polynucleotides encoding such proteins (such as, for example, in gene therapies or vectors suitable for introduction of DNA).

Research Uses and Utilities The polynucleotides provided by the present invention can be used by the research community for various purposes. The polynucleotides can be used to express recombinant protein for analysis, characterization or therapeutic use; as markers for tissues in which the corresponding protein is preferentially expressed (either constitutively or at a particular stage of tissue differentiation or development or in disease states); as molecular weight markers on Southern gels; as chromosome markers or tags (when labeled) to identify chromosomes or to map related gene positions; to compare with endogenous DNA sequences in patients to identify potential genetic disorders; as probes to hybridize and thus discover novel, related DNA sequences; as a source of information to derive PCR primers for genetic fingerprinting; as a probe to "subtract-out" known sequences in the process of discovering other novel polynucleotides; for selecting and making oligomers for attachment to a "gene chip" or other support, including for examination of expression patterns; to raise anti-protein antibodies using DNA immunization techniques; and as an antigen to raise anti-DNA antibodies or elicit another immune response. Where the polynucleotide encodes a protein which binds or potentially binds to another protein (such as, for example, in a receptor-ligand interaction), the polynucleotide can also be used in interaction trap assays (such as, for example, that described in Gyuris et al., Cell 75:791-803 (1993)) to identify polynucleotides encoding the other protein with which binding occurs or to identify inhibitors of the binding interaction. The proteins provided by the present invention can similarly be used in assay to determine biological activity, including in a panel of multiple proteins for high- throughput screening; to raise antibodies or to elicit another immune response; as a reagent (including the labeled reagent) in assays designed to quantitatively determine levels of the protein (or its receptor) in biological fluids; as markers for tissues in which the corresponding protein is preferentially expressed (either constitutively or at a particular stage of tissue differentiation or development or in a disease state); and, of course, to isolate correlative receptors or ligands. Where the protein binds or potentially binds to another protein (such as, for example, in a receptor-ligand interaction), the protein can be used to identify the other protein with which binding occurs or to identify inhibitors of the binding interaction. Proteins involved in these binding interactions can also be used to screen for peptide or small molecule inhibitors or agonists of the binding interaction.

Any or all of these research utilities are capable of being developed into reagent grade or kit format for commercialization as research products.

Methods for performing the uses listed above are well known to those skilled in the art. References disclosing such methods include without limitation "Molecular Cloning: A Laboratory Manual", 2d ed., Cold Spring Harbor Laboratory Press, Sambrook, J., E.F. Fritsch and T. Maniatis eds., 1989, and "Methods in Enzymology: Guide to Molecular Cloning Techniques", Academic Press, Berger, S.L. and A.R. Kimmel eds., 1987.

Nutritional Uses

Polynucleotides and proteins of the present invention can also be used as nutritional sources or supplements. Such uses include without limitation use as a protein or amino acid supplement, use as a carbon source, use as a nitrogen source and use as a source of carbohydrate. In such cases the protein or polynucleotide of the invention can be added to the feed of a particular organism or can be administered as a separate solid or liquid preparation, such as in the form of powder, pills, solutions, suspensions or capsules. In the case of microorganisms, the protein or polynucleotide of the invention can be added to the medium in or on which the microorganism is cultured.

Cytokine and Cell Proliferation /Differentiation Activity

A protein of the present invention may exhibit cytokine, cell proliferation (either inducing or inhibiting) or cell differentiation (either inducing or iriWbiting) activity or may induce production of other cytokines in certain cell populations. Many protein factors discovered to date, including all known cytokines, have exhibited activity in one or more factor dependent cell proliferation assays, and hence the assays serve as a convenient confirmation of cytokine activity. The activity of a protein of the present invention is evidenced by any one of a number of routine factor dependent cell proliferation assays for cell lines including, without limitation, 32D, DA2, DA1G, T10, B9, B9/11, BaF3, MC9/G, M+ (preB M+), 2E8, RB5, DAI, 123, T1165, HT2, CTLL2, TF-1, Mo7e and CMK.

The activity of a protein of the invention may, among other means, be measured by the following methods:

Assays for T-cell or thymocyte proliferation include without limitation those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley- Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1-3.19; Chapter 7, Immimologic studies in Humans); Takai et al, J. Immunol. 137:3494-3500, 1986; Bertagnolli et al., J. Immunol. 145:1706-1712, 1990; Bertagnolli et al., Cellular Immunology 133:327-341, 1991; Bertagnolli, et al, J. Immunol. 149:3778-3783, 1992; Bowman et al., J. Immunol. 152: 1756-1761, 1994.

Assays for cytokine production and /or proliferation of spleen cells, lymph node cells or thymocytes include, without limitation, those described in: Polyclonal T cell stimulation, Kruisbeek, A.M. and Shevach, E.M. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 3.12.1-3.12.14, John Wiley and Sons, Toronto. 1994; and Measurement of mouse and human Interferon γ, Schreiber, R.D. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.8.1-6.8.8, John Wiley and Sons, Toronto. 1994. Assays for proliferation and differentiation of hematopoietic and lymphopoietic cells include, without limitation, those described in: Measurement of Human and Murine Interleukin 2 and Interleukin 4, Bottomly, K., Davis, L.S. and Lipsky, P.E. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.3.1-6.3.12, John Wiley and Sons, Toronto. 1991; deVries et al., J. Exp. Med. 173:1205-1211, 1991; Moreau et al., Nature 336:690-692, 1988; Greenberger et al., Proc. Natl. Acad. Sci. U.S.A. 80:2931-2938, 1983; Measurement of mouse and human interleukin 6 - Nordan, R. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.6.1-6.6.5, John Wiley and Sons, Toronto. 1991; Smith et al., Proc. Natl. Acad. Sci. U.S.A. 83:1857-1861, 1986; Measurement of human Interleukin 11 - Bennett, F., Giannotti, J., Clark, S.C. and Turner, K. J. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.15.1 John Wiley and Sons, Toronto. 1991;

Measurement of mouse and human Interleukin 9 - Ciarletta, A., Giannotti, J., Clark, S.C. and Turner, K.J. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.13.1, John Wiley and Sons, Toronto. 1991. Assays for T-cell clone responses to antigens (which will identify, among others, proteins that affect APC-T cell interactions as well as direct T-cell effects by measuring proliferation and cytokine production) include, without limitation, those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function; Chapter 6, Cytokines and their cellular receptors; Chapter 7, Immunologic studies in Humans); Weinberger et al., Proc. Natl. Acad. Sci. USA 77:6091-6095, 1980; Weinberger et al., Eur. J. Immun. 11:405-411, 1981; Takai et al., J. Immunol. 137:3494-3500, 1986; Takai et al., J. Immunol. 140:508-512, 1988.

Immune Stimulating or Suppressing Activity

A protein of the present invention may also exhibit immune stimulating or immune suppressing activity, including without limitation the activities for which assays are described herein. A protein may be useful in the treatment of various immune deficiencies and disorders (including severe combined immunodeficiency (SOD)), e.g., in regulating (up or down) growth and proliferation of T and /or B lymphocytes, as well as effecting the cytolytic activity of NK cells and other cell populations. These immune deficiencies may be genetic or be caused by viral (e.g., HIV) as well as bacterial or fungal infections, or may result from autoimmune disorders. More specifically, infectious diseases causes by viral, bacterial, fungal or other infection may be treatable using a protein of the present invention, including infections by HIV, hepatitis viruses, herpesviruses, mycobacteria, Leishmania spp., malaria spp. and various fungal infections such as candidiasis. Of course, in this regard, a protein of the present invention may also be useful where a boost to the immune system generally may be desirable, i.e., in the treatment of cancer.

Autoimmune disorders which may be treated using a protein of the present invention include, for example, connective tissue disease, multiple sclerosis, systemic lupus erythematosus, rheumatoid arthritis, autoimmune pulmonary inflammation, Guillain-Barre syndrome, autoimmune thyroiditis, insulin dependent diabetes mellitis, myasthenia gravis, graft-versus-host disease and autoimmune inflammatory eye disease. Such a protein of the present invention may also to be useful in the treatment of allergic reactions and conditions, such as asthma (particularly allergic asthma) or other respiratory problems. Other conditions, in which immune suppression is desired (including, for example, organ transplantation), may also be treatable using a protein of the present invention.

Using the proteins of the invention it may also be possible to immune responses, in a number of ways. Down regulation may be in the form of inhibiting or blocking an immune response already in progress or may involve preventing the induction of an immune response. The functions of activated T cells may be inhibited by suppressing T cell responses or by inducing specific tolerance in T cells, or both. Immunosuppression of T cell responses is generally an active, non-antigen-specific, process which requires continuous exposure of the T cells to the suppressive agent. Tolerance, which involves inducing non-responsiveness or anergy in T cells, is distinguishable from immunosuppression in that it is generally antigen-specific and persists after exposure to the tolerizing agent has ceased. Operationally, tolerance can be demonstrated by the lack of a T cell response upon reexposure to specific antigen in the absence of the tolerizing agent. Down regulating or preventing one or more antigen functions (including without limitation B lymphocyte antigen functions (such as , for example, B7)), e.g., preventing high level lymphokine synthesis by activated T cells, will be useful in situations of tissue, skin and organ transplantation and in graft-versus-host disease (GVHD). For example, blockage of T cell function should result in reduced tissue destruction in tissue transplantation. Typically, in tissue transplants, rejection of the transplant is initiated through its recognition as foreign by T cells, followed by an immune reaction that destroys the transplant. The administration of a molecule which inhibits or blocks interaction of a B7 lymphocyte antigen with its natural ligand(s) on immune cells (such as a soluble, monomeric form of a peptide having B7-2 activity alone or in conjunction with a monomeric form of a peptide having an activity of another B lymphocyte antigen (e.g., B7-

1, B7-3) or blocking antibody), prior to transplantation can lead to the binding of the molecule to the natural ligand(s) on the immune cells without transmitting the corresponding costimulatory signal. Blocking B lymphocyte antigen function in this matter prevents cytokine synthesis by immune cells, such as T cells, and thus acts as an immunosuppressant. Moreover, the lack of costimulation may also be sufficient to anergize the T cells, thereby inducing tolerance in a subject. Induction of long-term tolerance by B lymphocyte antigen-blocking reagents may avoid the necessity of repeated administration of these blocking reagents. To achieve sufficient immunosuppression or tolerance in a subject, it may also be necessary to block the function of a combination of

B lymphocyte antigens.

The efficacy of particular blocking reagents in preventing organ transplant rejection or GVHD can be assessed using animal models that are predictive of efficacy in humans. Examples of appropriate systems which can be used include allogeneic cardiac grafts in rats and xenogeneic pancreatic islet cell grafts in mice, both of which have been used to examine the immunosuppressive effects of CTLA4Ig fusion proteins in vivo as described in Lenschow et al., Science 257:789-792 (1992) and Turka et al, Proc. Natl. Acad.

Sci USA, 89:11102-11105 (1992). In addition, murine models of GVHD (see Paul ed., Fundamental Immunology, Raven Press, New York, 1989, pp. 846-847) can be used to determine the effect of blocking B lymphocyte antigen function in vivo on the development of that disease.

Blocking antigen function may also be therapeutically useful for treating autoimmune diseases. Many autoimmune disorders are the result of inappropriate activation of T cells that are reactive against self tissue and which promote the production of cytokines and autoantibodies involved in the pathology of the diseases. Preventing the activation of autoreactive T cells may reduce or eliminate disease symptoms.

Administration of reagents which block costimulation of T cells by disrupting receptor igand interactions of B lymphocyte antigens can be used to inhibit T cell activation and prevent production of autoantibodies or T cell-derived cytokines which may be involved in the disease process. Additionally, blocking reagents may induce antigen-specific tolerance of autoreactive T cells which could lead to long-term relief from the disease. The efficacy of blocking reagents in preventing or alleviating autoimmune disorders can be determined using a number of well-characterized animal models of human autoimmune diseases. Examples include murine experimental autoimmune encephalitis, systemic lupus erythmatosis in MRL/lpr/lpr mice or NZB hybrid mice, murine autoimmune collagen arthritis, diabetes mellitus in NOD mice and BB rats, and murine experimental myasthenia gravis (see Paul ed., Fundamental Immunology, Raven Press, New York, 1989, pp. 840-856). Upregulation of an antigen function (preferably a B lymphocyte antigen function), as a means of up regulating immune responses, may also be useful in therapy. Upregulation of immune responses may be in the form of enhancing an existing immune response or eliciting an initial immune response. For example, enhancing an immune response through stimulating B lymphocyte antigen function may be useful in cases of viral infection. In addition, systemic viral diseases such as influenza, the common cold, and encephalitis might be alleviated by the administration of stimulatory forms of B lymphocyte antigens systemically.

Alternatively, anti-viral immune responses may be enhanced in an infected patient by removing T cells from the patient, costimulating the T cells in vitro with viral antigen- pulsed APCs either expressing a peptide of the present invention or together with a stimulatory form of a soluble peptide of the present invention and reintroducing the in vitro activated T cells into the patient. Another method of enhancing anti-viral immune responses would be to isolate infected cells from a patient, transfect them with a nucleic acid encoding a protein of the present invention as described herein such that the cells express all or a portion of the protein on their surface, and reintroduce the transfected cells into the patient. The infected cells would now be capable of delivering a costimulatory signal to, and thereby activate, T cells in vivo.

In another application, up regulation or enhancement of antigen function (preferably B lymphocyte antigen function) may be useful in the induction of tumor immunity. Tumor cells (e.g., sarcoma, melanoma, lymphoma, leukemia, neuroblastoma, carcinoma) transfected with a nucleic acid encoding at least one peptide of the present invention can be administered to a subject to overcome tumor-specific tolerance in the subject. If desired, the tumor cell can be transfected to express a combination of peptides. For example, tumor cells obtained from a patient can be transfected ex vivo with an expression vector directing the expression of a peptide having B7-2-like activity alone, or in conjunction with a peptide having B7-l-like activity and /or B7-3-like activity. The transfected umor cells are returned to the patient to result in expression of the peptides on the surface of the transfected cell. Alternatively, gene therapy techniques can be used to target a tumor cell for transfection in vivo.

The presence of the peptide of the present invention having the activity of a B lymphocyte antigen(s) on the surface of the tumor cell provides the necessary costimulation signal to T cells to induce a T cell mediated immune response against the transfected tumor cells. In addition, tumor cells which lack MHC class I or MHC class II molecules, or which fail to reexpress sufficient amounts of MHC class I or MHC class II molecules, can be transfected with nucleic acid encoding all or a portion of (e.g., a cytoplasmic-domain truncated portion) of an MHC class I a chain protein and β₂ microglobulin protein or an MHC class II α chain protein and an MHC class II β chain protein to thereby express MHC class I or MHC class II proteins on the cell surface. Expression of the appropriate class I or class II MHC in conjunction with a peptide having the activity of a B lymphocyte antigen (e.g., B7-1, B7-2, B7-3) induces a T cell mediated immune response against the transfected tumor cell. Optionally, a gene encoding an antisense construct which blocks expression of an MHC class II associated protein, such as the invariant chain, can also be cotransfected with a DNA encoding a peptide having the activity of a B lymphocyte antigen to promote presentation of tumor associated antigens and induce tumor specific immunity. Thus, the induction of a T cell mediated immune response in a human subject may be sufficient to overcome tumor-specific tolerance in the subject. The activity of a protein of the invention may, among other means, be measured by the following methods:

Suitable assays for thymocyte or splenocyte cytotoxicity include, without limitation, those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1- 3.19; Chapter 7, Immunologic studies in Humans); Herrmann et al., Proc. Natl. Acad. Sci. USA 78:2488-2492, 1981; Herrmann et al., J. Immunol. 128:1968-1974, 1982; Handa et al., J. Immunol. 135:1564-1572, 1985; Takai et al., J. Immunol. 137:3494-3500, 1986; Takai et al., J. Immunol. 140:508-512, 1988; Herrmann et al., Proc. Natl. Acad. Sci. USA 78:2488-2492, 1981; Herrmann et al., J. Immunol. 128:1968-1974, 1982; Handa et al., J. Immunol.

135:1564-1572, 1985; Takai et al, J. Immunol. 137:3494-3500, 1986; Bowmanet al., J.

Virology 61:1992-1998; Takai et al., J. Immunol. 140:508-512, 1988; Bertagnolli et al.,

Cellular Immunology 133:327-341, 1991; Brown et al., J. Immunol. 153:3079-3092, 1994.

Assays for T-cell-dependent immunoglobulin responses and isotype switching (which will identify, among others, proteins that modulate T-cell dependent antibody responses and that affect Thl/Th2 profiles) include, without limitation, those described in: Maliszewski, J. Immunol. 144:3028-3033, 1990; and Assays for B cell function: In vitro antibody production, Mond, J.J. and Brunswick, M. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 3.8.1-3.8.16, John Wiley and Sons, Toronto. 1994. Mixed lymphocyte reaction (MLR) assays (which will identify, among others, proteins that generate predominantly Thl and CTL responses) include, without limitation, those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley- Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1-3.19; Chapter 7, Immunologic studies in Humans); Takai et al., J. Immunol. 137:3494-3500, 1986; Takai et al., J. Immunol. 140:508-512, 1988; Bertagnolli et al., J. Immunol. 149:3778-3783, 1992.

Dendritic cell-dependent assays (which will identify, among others, proteins expressed by dendritic cells that activate naive T-cells) include, without limitation, those described in: Guery et al., J. Immunol. 134:536-544, 1995; Inaba et al., Journal of Experimental Medicine 173:549-559, 1991; Macatonia et al., Journal of Immunology 154:5071-5079, 1995; Porgador et al., Journal of Experimental Medicine 182:255-260, 1995; Nair et al., Journal of Virology 67:4062-4069, 1993; Huang et al., Science 264:961-965, 1994; Macatonia et al., Journal of Experimental Medicine 169:1255-1264, 1989; Bhardwaj et al., Journal of Clinical Investigation 94:797-807, 1994; and Inaba et al., Journal of Experimental Medicine 172:631-640, 1990.

Assays for lymphocyte survival /apoptosis (which will identify, among others, proteins that prevent apoptosis after superantigen induction and proteins that regulate lymphocyte homeostasis) include, without limitation, those described in: Darzynkiewicz et al., Cytometry 13:795-808, 1992; Gorczyca et al., Leukemia 7:659-670, 1993; Gorczyca et al., Cancer Research 53:1945-1951, 1993; Itoh et al., Cell 66:233-243, 1991; Zacharchuk, Journal of Immunology 145:4037-4045, 1990; Zamai et al., Cytometry 14:891-897, 1993; Gorczyca et al., International Journal of Oncology 1:639-648, 1992.

Assays for proteins that influence early steps of T-cell commitment and development include, without limitation, those described in: Antica et al., Blood

84:111-117, 1994; Fine et al., Cellular Immunology 155:111-122, 1994; Galy et al., Blood 85:2770-2778, 1995; Toki et al., Proc. Nat. Acad Sci. USA 88:7548-7551, 1991.

Hematopoiesis Regulating Activity A protein of the present invention may be useful in regulation of hematopoiesis and, consequently, in the treatment of myeloid or lymphoid cell deficiencies. Even marginal biological activity in support of colony forming cells or of factor-dependent cell lines indicates involvement in regulating hematopoiesis, e.g. in supporting the growth and proliferation of erythroid progenitor cells alone or in combination with other cytokines, thereby indicating utility, for example, in treating various anemias or for use in conjunction with irradiation/chemotherapy to stimulate the production of erythroid precursors and/or erythroid cells; in supporting the growth and proliferation of myeloid cells such as granulocytes and monocytes/macrophages (i.e., traditional CSF activity) useful, for example, in conjunction with chemotherapy to prevent or treat consequent myelo-suppression; in supporting the growth and proliferation of megakaryocytes and consequently of platelets thereby allowing prevention or treatment of various platelet disorders such as thrombocytopenia, and generally for use in place of or complimentary to platelet transfusions; and/or in supporting the growth and proliferation of hematopoietic stem cells which are capable of maturing to any and all of the above- mentioned hematopoietic cells and therefore find therapeutic utility in various stem cell disorders (such as those usually treated with transplantation, including, without limitation, aplastic anemia and paroxysmal nocturnal hemoglobinuria), as well as in repopulating the stem cell compartment post irradiation/chemotherapy, either in-vivo or ex-vivo (i.e., in conjunction with bone marrow transplantation or with peripheral progenitor cell transplantation (homologous or heterologous)) as normal cells or genetically manipulated for gene therapy.

The activity of a protein of the invention may, among other means, be measured by the following methods: Suitable assays for proliferation and differentiation of various hematopoietic lines are cited above.

Assays for embryonic stem cell differentiation (which will identify, among others, proteins that influence embryonic differentiation hematopoiesis) include, without limitation, those described in: Johansson et al. Cellular Biology 15:141-151, 1995; Keller et al., Molecular and Cellular Biology 13:473-486, 1993; McClanahan et al., Blood 81:2903-2915, 1993.

Assays for stem cell survival and differentiation (which will identify, among others, proteins that regulate lympho-hematopoiesis) include, without limitation, those described in: Methylcellulose colony forming assays, Freshney, M.G. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 265-268, Wiley-Liss, Inc., New York,

NY. 1994; Hirayama et al., Proc. Natl. Acad. Sci. USA 89:5907-5911, 1992; Primitive hematopoietic colony forming cells with high proliferative potential, McNiece, I.K. and Briddell, R.A. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 23-39, Wiley-Liss, Inc., New York, NY. 1994; Neben et al., Experimental Hematology 22:353-359, 1994; Cobblestone area forming cell assay, Ploemacher, R.E. In Culture of Hematopoietic

Cells. R.I. Freshney, et al. eds. Vol pp. 1-21, Wiley-Liss, Inc., New York, NY. 1994; Long term bone marrow cultures in the presence of stromal cells, Spooncer, E., Dexter, M. and Allen, T. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 163-179, Wiley-Liss, Inc., New York, NY. 1994; Long term culture initiating cell assay, Sutherland, HJ. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 139-162, Wiley-Liss, Inc., New York, NY. 1994.

Tissue Growth Activity A protein of the present invention also may have utility in compositions used for bone, cartilage, tendon, ligament and /or nerve tissue growth or regeneration, as well as for wound healing and tissue repair and replacement, and in the treatment of burns, incisions and ulcers.

A protein of the present invention, which induces cartilage and /or bone growth in circumstances where bone is not normally formed, has application in the healing of bone fractures and cartilage damage or defects in humans and other animals. Such a preparation employing a protein of the invention may have prophylactic use in closed as well as open fracture reduction and also in the improved fixation of artificial joints. De novo bone formation induced by an osteogenic agent contributes to the repair of congenital, trauma induced, or oncologic resection induced craniof acial defects, and also is useful in cosmetic plastic surgery.

A protein of this invention may also be used in the treatment of periodontal disease, and in other tooth repair processes. Such agents may provide an environment to attract bone-forming cells, stimulate growth of bone-forming cells or induce differentiation of progenitors of bone-forming cells. A protein of the invention may also be useful in the treatment of osteoporosis or osteoarthritis, such as through stimulation of bone and /or cartilage repair or by blocking inflammation or processes of tissue destruction (collagenase activity, osteoclast activity, etc.) mediated by inflammatory processes. Another category of tissue regeneration activity that may be attributable to the protein of the present invention is tendon/ligament formation. A protein of the present invention, which induces tendon/ligament-like tissue or other tissue formation in circumstances where such tissue is not normally formed, has application in the healing of tendon or ligament tears, deformities and other tendon or ligament defects in humans and other animals. Such a preparation employing a tendon/ligament-like tissue inducing protein may have prophylactic use in preventing damage to tendon or ligament tissue, as well as use in the improved fixation of tendon or ligament to bone or other tissues, and in repairing defects to tendon or ligament tissue. De novo tendon/ligament-like tissue formation induced by a composition of the present invention contributes to the repair of congenital, trauma induced, or other tendon or ligament defects of other origin, and is also useful in cosmetic plastic surgery for attachment or repair of tendons or ligaments. The compositions of the present invention may provide an environment to attract tendon- or ligament-forming cells, stimulate growth of tendon- or ligament-forming cells, induce differentiation of progenitors of tendon- or ligament-forming cells, or induce growth of tendon/ligament cells or progenitors ex vivo for return in vivo to effect tissue repair. The compositions of the invention may also be useful in the treatment of tendinitis, carpal tunnel syndrome and other tendon or ligament defects. The compositions may also include an appropriate matrix and /or sequestering agent as a carrier as is well known in the art.

The protein of the present invention may also be useful for proliferation of neural cells and for regeneration of nerve and brain tissue, i.e. for the treatment of central and peripheral nervous system diseases and neuropathies, as well as mechanical and traumatic disorders, which involve degeneration, death or trauma to neural cells or nerve tissue. More specifically, a protein may be used in the treatment of diseases of the peripheral nervous system, such as peripheral nerve injuries, peripheral neuropathy and localized neuropathies, and central nervous system diseases, such as Alzheimer's, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and Shy-Drager syndrome. Further conditions which may be treated in accordance with the present invention include mechanical and traumatic disorders, such as spinal cord disorders, head trauma and cerebrovascular diseases such as stroke. Peripheral neuropathies resulting from chemotherapy or other medical therapies may also be treatable using a protein of the invention.

Proteins of the invention may also be useful to promote better or faster closure of non-healing wounds, including without limitation pressure ulcers, ulcers associated with vascular insufficiency, surgical and traumatic wounds, and the like.

It is expected that a protein of the present invention may also exhibit activity for generation or regeneration of other tissues, such as organs (including, for example, pancreas, liver, intestine, kidney, skin, endothelium), muscle (smooth, skeletal or cardiac) and vascular (including vascular endothelium) tissue, or for promoting the growth of cells comprising such tissues. Part of the desired effects may be by inhibition or modulation of fibrotic scarring to allow normal tissue to regenerate. A protein of the invention may also exhibit angioge ic activity. A protein of the present invention may also be useful for gut protection or regeneration and treatment of lung or liver fibrosis, reperfusion injury in various tissues, and conditions resulting from systemic cytokine damage.

A protein of the present invention may also be useful for promoting or inhibiting differentiation of tissues described above from precursor tissues or cells; or for inhibiting the growth of tissues described above.

Assays for tissue generation activity include, without limitation, those described in: International Patent Publication No. W095/ 16035 (bone, cartilage, tendon); International Patent Publication No. WO95/05846 (nerve, neuronal); International Patent Publication No. WO91/07491 (skin, endothelium ).

Assays for wound healing activity include, without limitation, those described in: Winter, Epidermal Wound Healing, pps. 71-112 (Maibach, HI and Rovee, DT, eds.), Year Book Medical Publishers, Inc., Chicago, as modified by Eaglstein and Mertz, J. Invest. Dermatol 71:382-84 (1978).

Activin /Inhibin Activity

A protein of the present invention may also exhibit activin- or inhibin-related activities. Inhibins are characterized by their ability to inhibit the release of follicle stimulating hormone (FSH), while activins and are characterized by their ability to stimulate the release of follicle stimulating hormone (FSH). Thus, a protein of the present invention, alone or in heterodimers with a member of the inhibin a family, may be useful as a contraceptive based on the ability of inhibins to decrease fertility in female mammals and decrease spermatogenesis in male mammals. Administration of sufficient amounts of other inhibins can induce infertility in these mammals. Alternatively, the protein of the invention, as a homodimer or as a heterodimer with other protein subunits of the inhibin- β group, may be useful as a fertility inducing therapeutic, based upon the ability of activin molecules in stimulating FSH release from cells of the anterior pituitary. See, for example, United States Patent 4,798,885. A protein of the invention may also be useful for advancement of the onset of fertility in sexually immature mammals, so as to increase the lifetime reproductive performance of domestic animals such as cows, sheep and pigs.

The activity of a protein of the invention may, among other means, be measured by the following methods: Assays for activm/inhibin activity include, without limitation, those described in: Vale et al., Endocrinology 91:562-572, 1972; Ling et al., Nature 321:779-782, 1986; Vale et al., Nature 321:776-779, 1986; Mason et al., Nature 318:659-663, 1985; Forage et al., Proc. Natl. Acad. Sci. USA 83:3091-3095, 1986.

Chemotactic/Chemokinetic Activity

A protein of the present invention may have chemotactic or chemokinetic activity (e.g., act as a chemokine) for mammalian cells, including, for example, monocytes, fibroblasts, neutrophils, T-cells, mast cells, eosinophils, epithelial and /or endothelial cells. Chemotactic and chemokinetic proteins can be used to mobilize or attract a desired cell population to a desired site of action. Chemotactic or chemokinetic proteins provide particular advantages in treatment of wounds and other trauma to tissues, as well as in treatment of localized infections. For example, attraction of lymphocytes, monocytes or neutrophils to tumors or sites of infection may result in improved immune responses against the tumor or infecting agent.

A protein or peptide has chemotactic activity for a particular cell population if it can stimulate, directly or indirectly, the directed orientation or movement of such cell population. Preferably, the protein or peptide has the ability to directly stimulate directed movement of cells. Whether a particular protein has chemotactic activity for a population of cells can be readily determined by employing such protein or peptide in any known assay for cell chemotaxis.

Assays for chemotactic activity (which will identify proteins that induce or prevent chemotaxis) consist of assays that measure the ability of a protein to induce the migration of cells across a membrane as well as the ability of a protein to induce the adhesion of one cell population to another cell population. Suitable assays for movement and adhesion include, without limitation, those described in: Current Protocols in Immunology, Ed by J.E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W.Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 6.12, Measurement of alpha and beta Chemokines 6.12.1-6.12.28; Taub et al. J. Clin. Invest. 95:1370-1376, 1995; Lind et al. APMIS 103:140-146, 1995; Muller et al Eur. J. Immunol. 25: 1744-1748; Gruber et al. J. of Immunol. 152:5860-5867, 1994; Johnston et al. J. of Immunol. 153: 1762-1768, 1994. Hemostatic and Thrombolytic Activity

A protein of the invention may also exhibit hemostatic or thrombolytic activity. As a result, such a protein is expected to be useful in treatment of various coagulation disorders (including hereditary disorders, such as hemophilias) or to enhance coagulation and other hemostatic events in treating wounds resulting from trauma, surgery or other causes. A protein of the invention may also be useful for dissolving or inhibiting formation of thromboses and for treatment and prevention of conditions resulting therefrom (such as, for example, infarction of cardiac and central nervous system vessels (e.g., stroke). The activity of a protein of the invention may, among other means, be measured by the following methods:

Assay for hemostatic and thrombolytic activity include, without limitation, those described in: Linet et al., J. Clin. Pharmacol. 26:131-140, 1986; Burdick et al, Thrombosis Res.45:413-419, 1987; Humphrey et al., Fibrinolysis 5:71-79 (1991); Schaub, Prostaglandins 35:467-474, 1988.

Receptor/Ligand Activity

A protein of the present invention may also demonstrate activity as receptors, receptor ligands or inhibitors or agonists of receptor /ligand interactions. Examples of such receptors and ligands include, without limitation, cytokine receptors and their ligands, receptor kinases and their ligands, receptor phosphatases and their ligands, receptors involved in cell-cell interactions and their ligands (including without limitation, cellular adhesion molecules (such as selectins, integrins and their ligands) and receptor /ligand pairs involved in antigen presentation, antigen recognition and development of cellular and humoral immune responses). Receptors and ligands are also useful for screening of potential peptide or small molecule inhibitors of the relevant receptor/ligand interaction. A protein of the present invention (including, without limitation, fragments of receptors and ligands) may themselves be useful as inhibitors of receptor/ligand interactions. The activity of a protein of the invention may, among other means, be measured by the following methods:

Suitable assays for receptor-ligand activity include without limitation those described in:Current Protocols in Immunology, Ed by J.E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W.Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 7.28, Measurement of Cellular Adhesion under static conditions 7.28.1-7.28.22), Takai et al., Proc. Natl. Acad. Sci. USA 84:6864-6868, 1987; Bierer et al., J. Exp. Med. 168:1145-1156, 1988; Rosenstein et al., J. Exp. Med. 169:149-160 1989; Stoltenborg et al, J. Immunol. Methods 175:59-68, 1994; Stitt et al., Cell 80:661-670, 1995.

Anti-Inflammatory Activity

Proteins of the present invention may also exhibit anti-inflammatory activity. The anti-inflammatory activity may be achieved by providing a stimulus to cells involved in the inflammatory response, by inhibiting or promoting cell-cell interactions (such as, for example, cell adhesion), by inhibiting or promoting chemotaxis of cells involved in the inflammatory process, inhibiting or promoting cell extravasation, or by stimulating or suppressing production of other factors which more directly inhibit or promote an inflammatory response. Proteins exhibiting such activities can be used to treat inflammatory conditions including chronic or acute conditions), including without limitation inflammation associated with infection (such as septic shock, sepsis or systemic inflammatory response syndrome (SIRS)), ischemia-reperfusion injury, endotoxin lethality, arthritis, complement-mediated hyperacute rejection, nephritis, cytokine or chemokine-induced lung injury, inflammatory bowel disease, Crohn's disease or resulting from over production of cytokines such as TNF or IL-1. Proteins of the invention may also be useful to treat anaphylaxis and hypersensitivity to an antigenic substance or material.

Cadherin/Tumor Invasion Suppressor Activity

Cadherins are calcium-dependent adhesion molecules that appear to play major roles during development, particularly in defining specific cell types. Loss or alteration of normal cadherin expression can lead to changes in cell adhesion properties linked to tumor growth and metastasis. Cadherin malfunction is also implicated in other human diseases, such as pemphigus vulgaris and pemphigus foliaceus (auto-immune blistering skin diseases), Crohn's disease, and some developmental abnormalities. The cadherin superfamily includes well over forty members, each with a distinct pattern of expression. All members of the superfamily have in common conserved extracellular repeats (cadherin domains), but structural differences are found in other parts of the molecule. The cadherin domains bind calcium to form their tertiary structure and thus calcium is required to mediate their adhesion. Only a few amino acids in the first cadherin domain provide the basis for homophilic adhesion; modification of this recognition site can change the specificity of a cadherin so that instead of recognizing only itself, the mutant molecule can now also bind to a different cadherin. In addition, some cadherins engage in heterophilic adhesion with other cadherins. E-cadherin, one member of the cadherin superfamily, is expressed in epithelial cell types. Pathologically, if E-cadherin expression is lost in a tumor, the malignant cells become invasive and the cancer metastasizes. Transfection of cancer cell lines with polynucleotides expressing E-cadherin has reversed cancer-associated changes by returning altered cell shapes to normal, restoring cells' adhesiveness to each other and to their substrate, decreasing the cell growth rate, and drastically reducing anchorage- independent cell growth. Thus, reintroducing E-cadherin expression reverts carcinomas to a less advanced stage. It is likely that other cadherins have the same invasion suppressor role in carcinomas derived from other tissue types. Therefore, proteins of the present invention with cadherin activity, and polynucleotides of the present invention encoding such proteins, can be used to treat cancer. Introducing such proteins or polynucleotides into cancer cells can reduce or eliminate the cancerous changes observed in these cells by providing normal cadherin expression.

Cancer cells have also been shown to express cadherins of a different tissue type than their origin, thus allowing these cells to invade and metastasize in a different tissue in the body. Proteins of the present invention with cadherin activity, and polynucleotides of the present invention encoding such proteins, can be substituted in these cells for the inappropriately expressed cadherins, restoring normal cell adhesive properties and reducing or eliminating the tendency of the cells to metastasize.

Additionally, proteins of the present invention with cadherin activity, and polynucleotides of the present invention encoding such proteins, can used to generate antibodies recognizing and binding to cadherins. Such antibodies can be used to block the adhesion of inappropriately expressed tumor-cell cadherins, preventing the cells from forming a tumor elsewhere. Such an anti-cadherin antibody can also be used as a marker for the grade, pathological type, and prognosis of a cancer, i.e. the more progressed the cancer, the less cadherin expression there will be, and this decrease in cadherin expression can be detected by the use of a cadherin-binding antibody.

Fragments of proteins of the present invention with cadherin activity, preferably a polypeptide comprising a decapeptide of the cadherin recognition site, and polynucleotides of the present invention encoding such protein fragments, can also be used to block cadherin function by binding to cadherins and preventing them from binding in ways that produce undesirable effects. Additionally, fragments of proteins of the present invention with cadherin activity, preferably truncated soluble cadherin fragments which have been found to be stable in the circulation of cancer patients, and polynucleotides encoding such protein fragments, can be used to disturb proper cell-cell adhesion.

Assays for cadherin adhesive and invasive suppressor activity include, without limitation, those described in: Hortsch et al. J Biol Chem 270 (32): 18809-18817, 1995; Miyaki et al. Oncogene 11: 2547-2552, 1995; Ozawa et al. Cell 63: 1033-1038, 1990.

Tumor Inhibition Activity

In addition to the activities described above for immunological treatment or prevention of tumors, a protein of the invention may exhibit other anti-tumor activities. A protein may inhibit tumor growth directly or indirectly (such as, for example, via ADCC). A protein may exhibit its tumor inhibitory activity by acting on tumor tissue or tumor precursor tissue, by inhibiting formation of tissues necessary to support tumor growth (such as, for example, by inhibiting angiogenesis), by causing production of other factors, agents or cell types which inhibit tumor growth, or by suppressing, eliminating or inhibiting factors, agents or cell types which promote tumor growth.

Other Activities

A protein of the invention may also exhibit one or more of the following additional activities or effects: inhibiting the growth, infection or function of, or killing, infectious agents, including, without limitation, bacteria, viruses, fungi and other parasites; effecting (suppressing or enhancing) bodily characteristics, including, without limitation, height, weight, hair color, eye color, skin, fat to lean ratio or other tissue pigmentation, or organ or body part size or shape (such as, for example, breast augmentation or diminution, change in bone form or shape); effecting biorhythms or caricadic cycles or rhythms; effecting the fertility of male or female subjects; effecting the metabolism, catabolism, anabolism, processing, utilization, storage or elimination of dietary fat, lipid, protein, carbohydrate, vitamins, minerals, cofactors or other nutritional factors or component(s); effecting behavioral characteristics, including, without limitation, appetite, libido, stress, cognition (including cognitive disorders), depression (including depressive disorders) and violent behaviors; providing analgesic effects or other pain reducing effects; promoting differentiation and growth of embryonic stem cells in lineages other than hematopoietic lineages; hormonal or endocrine activity; in the case of enzymes, correcting deficiencies of the enzyme and treating deficiency-related diseases; treatment of hyperproliferative disorders (such as, for example, psoriasis); immunoglobulin-like activity (such as, for example, the ability to bind antigens or complement); and the ability to act as an antigen in a vaccine composition to raise an immune response against such protein or another material or entity which is cross-reactive with such protein.

ADMINISTRATION AND DOSING

A protein of the present invention (from whatever source derived, including without limitation from recombinant and non-recombinant sources) may be used in a pharmaceutical composition when combined with a pharmaceutically acceptable carrier. Such a composition may also contain (in addition to protein and a carrier) diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. The term "pharmaceutically acceptable" means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredient(s). The characteristics of the carrier will depend on the route of administration. The pharmaceutical composition of the invention may also contain cytokines, lymphokines, or other hematopoietic factors such as M-CSF, GM-CSF, TNF, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IFN, TNFO, TNF1, TNF2, G-CSF, Meg-CSF, thrombopoietin, stem cell factor, and erythropoietin. The pharmaceutical composition may further contain other agents which either enhance the activity of the protein or compliment its activity or use in treatment. Such additional factors and /or agents may be included in the pharmaceutical composition to produce a synergistic effect with protein of the invention, or to rni imize side effects. Conversely, protein of the present invention may be included in formulations of the particular cytokine, lymphokine, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti-inflammatory agent to minimize side effects of the cytokine, lymphokine, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti-inflammatory agent.

A protein of the present invention may be active in multimers (e.g., heterodimers or homodimers) or complexes with itself or other proteins. As a result, pharmaceutical compositions of the invention may comprise a protein of the invention in such multimeric or complexed form.

The pharmaceutical composition of the invention may be in the form of a complex of the protein(s) of present invention along with protein or peptide antigens. The protein and/or peptide antigen will deliver a stimulatory signal to both B and T lymphocytes. B lymphocytes will respond to antigen through their surface immunoglobulin receptor. T lymphocytes will respond to antigen through the T cell receptor (TCR) following presentation of the antigen by MHC proteins. MHC and structurally related proteins including those encoded by class I and class II MHC genes on host cells will serve to present the peptide antigen(s) to T lymphocytes. The antigen components could also be supplied as purified MHC-peptide complexes alone or with co-stimulatory molecules that can directly signal T cells. Alternatively antibodies able to bind surface immunolgobulin and other molecules on B cells as well as antibodies able to bind the TCR and other molecules on T cells can be combined with the pharmaceutical composition of the invention.

The pharmaceutical composition of the invention may be in the form of a liposome in which protein of the present invention is combined, in addition to other pharmaceutically acceptable carriers, with amphipathic agents such as lipids which exist in aggregated form as micelles, insoluble monolayers, liquid crystals, or lamellar layers in aqueous solution. Suitable lipids for liposomal formulation include, without limitation, monoglycerides, diglycerides, sulfatides, lysolecithin, phospholipids, saponin, bile acids, and the like. Preparation of such liposomal formulations is within the level of skill in the art, as disclosed, for example, in U.S. Patent No.4,235,871; U.S. Patent No. 4,501,728; U.S. Patent No. 4,837,028; and U.S. Patent No. 4,737,323, all of which are incorporated herein by reference.

As used herein, the term "therapeutically effective amount" means the total amount of each active component of the pharmaceutical composition or method that is sufficient to show a meaningful patient benefit, i.e., treatment, healing, prevention or amelioration of the relevant medical condition, or an increase in rate of treatment, healing, prevention or amelioration of such conditions. When applied to an individual active ingredient, administered alone, the term refers to that ingredient alone. When applied to a combination, the term refers to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously. In practicing the method of treatment or use of the present invention, a therapeutically effective amount of protein of the present invention is administered to a mammal having a condition to be treated. Protein of the present invention may be administered in accordance with the method of the invention either alone or in combination with other therapies such as treatments employing cytokines, lymphokines or other hematopoietic factors. When co-administered with one or more cytokines, lymphokines or other hematopoietic factors, protein of the present invention may be administered either simultaneously with the cytokine(s), lymphokine(s), other hematopoietic factor(s), thrombolytic or anti-thrombotic factors, or sequentially. If administered sequentially, the attending physician will decide on the appropriate sequence of administering protein of the present invention in combination with cytokine(s), lymphokine(s), other hematopoietic factor(s), thrombolytic or anti-thrombotic factors.

Administration of protein of the present invention used in the pharmaceutical composition or to practice the method of the present invention can be carried out in a variety of conventional ways, such as oral ingestion, inhalation, topical application or cutaneous, subcutaneous, intraperitoneal, parenteral or intravenous injection. Intravenous administration to the patient is preferred.

When a therapeutically effective amount of protein of the present invention is administered orally, protein of the present invention will be in the form of a tablet, capsule, powder, solution or elixir. When administered in tablet form, the pharmaceutical composition of the invention may additionally contain a solid carrier such as a gelatin or an adjuvant. The tablet, capsule, and powder contain from about 5 to 95% protein of the present invention, and preferably from about 25 to 90% protein of the present invention. When administered in liquid form, a liquid carrier such as water, petroleum, oils of animal or plant origin such as peanut oil, mineral oil, soybean oil, or sesame oil, or synthetic oils may be added. The liquid form of the pharmaceutical composition may further contain physiological saline solution, dextrose or other saccharide solution, or glycols such as ethylene glycol, propylene glycol or polyethylene glycol. When administered in liquid form, the pharmaceutical composition contains from about 0.5 to 90% by weight of protein of the present invention, and preferably from about 1 to 50% protein of the present invention.

When a therapeutically effective amount of protein of the present invention is administered by intravenous, cutaneous or subcutaneous injection, protein of the present invention will be in the form of a pyrogen-free, parenterally acceptable aqueous solution.

The preparation of such parenterally acceptable protein solutions, having due regard to pH, isotonicity, stability, and the like, is within the skill in the art. A preferred pharmaceutical composition for intravenous, cutaneous, or subcutaneous injection should contain, in addition to protein of the present invention, an isotonic vehicle such as Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, Lactated Ringer's Injection, or other vehicle as known in the art. The pharmaceutical composition of the present invention may also contain stabilizers, preservatives, buffers, antioxidants, or other additives known to those of skill in the art. The amount of protein of the present invention in the pharmaceutical composition of the present invention will depend upon the nature and severity of the condition being treated, and on the nature of prior treatments which the patient has undergone. Ultimately, the attending physician will decide the amount of protein of the present invention with which to treat each individual patient. Initially, the attending physician will administer low doses of protein of the present invention and observe the patient's response. Larger doses of protein of the present invention may be administered until the optimal therapeutic effect is obtained for the patient, and at that point the dosage is not increased further. It is contemplated that the various pharmaceutical compositions used to practice the method of the present invention should contain about 0.01 μg to about 100 mg (preferably about O.lng to about 10 mg, more preferably about 0.1 μg to about 1 mg) of protein of the present invention per kg body weight.

The duration of intravenous therapy using the pharmaceutical composition of the present invention will vary, depending on the severity of the disease being treated and the condition and potential idiosyncratic response of each individual patient. It is contemplated that the duration of each application of the protein of the present invention will be in the range of 12 to 24 hours of continuous intravenous administration. Ultimately the attending physician will decide on the appropriate duration of intravenous therapy using the pharmaceutical composition of the present invention.

Protein of the invention may also be used to immunize animals to obtain polyclonal and monoclonal antibodies which specifically react with the protein. Such antibodies may be obtained using either the entire protein or fragments thereof as an immunogen. The peptide immunogens additionally may contain a cysteine residue at the carboxyl terminus, and are conjugated to a hapten such as keyhole limpet hemocyanin (KLH). Methods for synthesizing such peptides are known in the art, for example, as in R.P. Merrifield, J. Amer.Chem.Soc. 85, 2149-2154 (1963); J.L. Krstenansky, et al, FEBS Lett.

211, 10 (1987). Monoclonal antibodies binding to the protein of the invention may be useful diagnostic agents for the immunodetection of the protein. Neutralizing monoclonal antibodies binding to the protein may also be useful therapeutics for both conditions associated with the protein and also in the treatment of some forms of cancer where abnormal expression of the protein is involved. In the case of cancerous cells or leukemic cells, neutralizing monoclonal antibodies against the protein may be useful in detecting and preventing the metastatic spread of the cancerous cells, which may be mediated by the protein. For compositions of the present invention which are useful for bone, cartilage, tendon or ligament regeneration, the therapeutic method includes administering the composition topically, systematically, or locally as an implant or device. When administered, the therapeutic composition for use in this invention is, of course, in a pyrogen-free, physiologically acceptable form. Further, the composition may desirably be encapsulated or injected in a viscous form for delivery to the site of bone, cartilage or tissue damage. Topical administration may be suitable for wound healing and tissue repair. Therapeutically useful agents other than a protein of the invention which may also optionally be included in the composition as described above, may alternatively or additionally, be administered simultaneously or sequentially with the composition in the methods of the invention. Preferably for bone and /or cartilage formation, the composition would include a matrix capable of delivering the protein-containing composition to the site of bone and/or cartilage damage, providing a structure for the developing bone and cartilage and optimally capable of being resorbed into the body. Such matrices may be formed of materials presently in use for other implanted medical applications.

The choice of matrix material is based on biocompatibility, biodegradability, mechanical properties, cosmetic appearance and interface properties. The particular application of the compositions will define the appropriate formulation. Potential matrices for the compositions may be biodegradable and chemically defined calcium sulfate, tricalciumphosphate, hydroxyapatite, polylactic acid, polyglycolic acid and polyanhydrides. Other potential materials are biodegradable and biologically well- defined, such as bone or dermal collagen. Further matrices are comprised of pure proteins or extracellular matrix components. Other potential matrices are nonbiodegradable and chemically defined, such as sintered hydroxapatite, bioglass, aluminates, or other ceramics. Matrices may be comprised of combinations of any of the above mentioned types of material, such as polylactic acid and hydroxyapatite or collagen and tricalciumphosphate. The bioceramics may be altered in composition, such as in calcium- aluminate-phosphate and processing to alter pore size, particle size, particle shape, and biodegradability. Presently preferred is a 50:50 (mole weight) copolymer of lactic acid and glycolic acid in the form of porous particles having diameters ranging from 150 to 800 microns.

In some applications, it will be useful to utilize a sequestering agent, such as carboxymethyl cellulose or autologous blood clot, to prevent the protein compositions from disassociating from the matrix.

A preferred family of sequestering agents is cellulosic materials such as alkylcelluloses (including hydroxyalkylcelluloses), including methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl- methylcellulose, and carboxymethylcellulose, the most preferred being cationic salts of carboxymethylcellulose (CMC). Other preferred sequestering agents include hyaluronic acid, sodium alginate, poly(ethylene glycol), polyoxyethylene oxide, carboxyvinyl polymer and poly(vinyl alcohol). The amount of sequestering agent useful herein is 0.5-20 wt%, preferably 1-10 wt% based on total formulation weight, which represents the amount necessary to prevent desorbtion of the protein from the polymer matrix and to provide appropriate handling of the composition, yet not so much that the progenitor cells are prevented from infiltrating the matrix, thereby providing the protein the opportunity to assist the osteogenic activity of the progenitor cells.

In further compositions, proteins of the invention may be combined with other agents beneficial to the treatment of the bone and /or cartilage defect, wound, or tissue in question. These agents include various growth factors such as epidermal growth factor

(EGF), platelet derived growth factor (PDGF), transforming growth factors (TGF-α and

TGF-β), and insulin-like growth factor (IGF).

The therapeutic compositions are also presently valuable for veterinary applications. Particularly domestic animals and thoroughbred horses, in addition to humans, are desired patients for such treatment with proteins of the present invention.

The dosage regimen of a protein-containing pharmaceutical composition to be used in tissue regeneration will be determined by the attending physician considering various factors which modify the action of the proteins, e.g., amount of tissue weight desired to be formed, the site of damage, the condition of the damaged tissue, the size of a wound, type of damaged tissue (e.g., bone), the patient's age, sex, and diet, the severity of any infection, time of administration and other clinical factors. The dosage may vary with the type of matrix used in the reconstitution and with inclusion of other proteins in the pharmaceutical composition. For example, the addition of other known growth factors, such as IGF I (insulin like growth factor I), to the final composition, may also effect the dosage. Progress can be monitored by periodic assessment of tissue/bone growth and/or repair, for example, X-rays, histomorphometric determinations and tetracycline labeling.

Polynucleotides of the present invention can also be used for gene therapy. Such polynucleotides can be introduced either in vivo or ex vivo into cells for expression in a mammalian subject. Polynucleotides of the invention may also be administered by other known methods for introduction of nucleic acid into a cell or organism (including, without limitation, in the form of viral vectors or naked DNA).

Cells may also be cultured ex vivo in the presence of proteins of the present invention in order to proliferate or to produce a desired effect on or activity in such cells. Treated cells can then be introduced in vivo for therapeutic purposes.

Patent and literature references cited herein are incorporated by reference as if fully set forth.

SEQUENCE LISTING

(1) GENERAL INFORMATION:

(i) APPLICANT: Jacobs, Kenneth McCoy, John M. LaVallie, Edward R. Racie, Lisa A. Merberg, David Treacy, Maurice Spaulding, Vikki Agostino, Michael J.

(ii) TITLE OF INVENTION: SECRETED PROTEINS AND POLYNUCLEOTIDES ENCODING THEM

(iii) NUMBER OF SEQUENCES: 30

(iv) CORRESPONDENCE ADDRESS:

(A) ADDRESSEE: Genetics Institute, Inc.

(B) STREET: 87 CambridgePark Drive

(C) CITY: Cambridge

(D) STATE: MA

(E) COUNTRY: U.S.A.

(F) ZIP: 02140

(v) COMPUTER READABLE FORM:

(A) MEDIUM TYPE: Floppy disk

(B) COMPUTER: IBM PC compatible

(C) OPERATING SYSTEM: PC-DOS/MS-DOS

(D) SOFTWARE: Patentin Release #1.0, Version #1.30

(vi) CURRENT APPLICATION DATA:

(A) APPLICATION NUMBER:

(B) FILING DATE:

(C) CLASSIFICATION:

(viii) ATTORNEY/AGENT INFORMATION:

(A) NAME: Sprunger, Suzanne A.

(B) REGISTRATION NUMBER: 41,323

(ix) TELECOMMUNICATION INFORMATION:

(A) TELEPHONE: (617) 498-8284

(B) TELEFAX: (617) 876-5851

(2) INFORMATION FOR SEQ ID NO : 1 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 886 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS : double

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 1 :

CGCCCAGGCT GCCAGACCGG AAGCGCTCCG CTGTACCTGG ATCCTGCTCC TCTGGGTTGA 60

AACCCGGGCG CCGCCAAGAT GCCGGCTTAC CACTCTTCTC TCATGGATCC TGATACCAAA 120

CTCATCGGAA ACATGGCACT GTTGCCTATC AGAAGTCAAT TCAAAGGACC TGCCCCCAGA 180

GAGACAAAAG ATACAGATAT TGTGGATGAA GCCATCTATT ACTTCAAGGC CAATGTCTTC 240

TTCAAAAACT ATGAAATTAA GAATGAAGCT GATAGGACCT TGATATATAT AACTCTCTAC 300

ATTTCTGAAT GTCTGAAGAA ACTGCAAAAG TGCAATTCCA AAAGCCAAGG TGAGAAAGAA 360

ATGTATACGC TGGGAATCAC TAATTTTCCC ATTCCTGGAG AGCCTGGTTT TCCACTTAAC 420

GCAATTTATG CCAAACCTGC AAACAAACAG GAAGATGAAG TGATGAGAGC CTATTTACAA 480

CAGCTAAGGC AAGAGACTGG ACTGAGACTT TGTGAGAAAG TTTTCGACCC TCAGAATGAT 540

AAACCCAGCA AGTGGTGGAC TTGCTTTGTG AAGAGACAGT TCATGAACAA GAGTCTTTCA 600

GGACCTGGAC AGTGAAGGGA GCCCGGGCAG CCACCGTCTC CAGAGCCCTG GGCAGCATTT 660

TCCAGCAAGA TGTACACAAT CTTTTGCCTT TATTTCGTAA AGTTTTATAC AGAAGAGAGA 720

AGAGCATGTC TTTACTTGAA AAACTCTTGA TCAAGAATTT GGGTGGGAGA AAAGAAAGTG 780

GGTTATCAAG GGTGATTTGA AATTTTCTGC AGCATTAAAG CTGGCGCTTA ATAAGAATAA 840

GTAATAATAA AGAAATTTCT AACAAAAAAA AAAAAAAAAA AAAAAA 886 (2) INFORMATION FOR SEQ ID NO : 2 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 178 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2:

Met Pro Ala Tyr His Ser Ser Leu Met Asp Pro Asp Thr Lys Leu lie 1 5 10 15 Gly Asn Met Ala Leu Leu Pro lie Arg Ser Gin Phe Lys Gly Pro Ala 20 25 30

Pro Arg Glu Thr Lys Asp Thr Asp lie Val Asp Glu Ala lie Tyr Tyr 35 40 45

Phe Lys Ala Asn Val Phe Phe Lys Asn Tyr Glu lie Lys Asn Glu Ala 50 55 60

Asp Arg Thr Leu lie Tyr lie Thr Leu Tyr lie Ser Glu Cys Leu Lys 65 70 75 80

Lys Leu Gin Lys Cys Asn Ser Lys Ser Gin Gly Glu Lys Glu Met Tyr 85 90 95

Thr Leu Gly lie Thr Asn Phe Pro lie Pro Gly Glu Pro Gly Phe Pro 100 105 110

Leu Asn Ala lie Tyr Ala Lys Pro Ala Asn Lys Gin Glu Asp Glu Val 115 120 125

Met Arg Ala Tyr Leu Gin Gin Leu Arg Gin Glu Thr Gly Leu Arg Leu 130 135 140

Cys Glu Lys Val Phe Asp Pro Gin Asn Asp Lys Pro Ser Lys Trp Trp 145 150 155 160

Thr Cys Phe Val Lys Arg Gin Phe Met Asn Lys Ser Leu Ser Gly Pro 165 170 175

Gly Gin

( 2 ) INFORMATION FOR SEQ ID NO : 3 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1537 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 3 :

GCGATGAACG TGACTTACTT ACTGTCAACC GGAAAATAAA AATATCTAAC CTTGAAAAGG 60

AACAAATGCT CACCTCTGAC TTTAAGAAAA ATACCAGACT ATTACCAAAA TTGAAGAAAA 120

TAGAAAATCA GGTAGCTATG TCATTTTATA AGCATCAGTC CTCACCAGAT TTGTCAAGTG 180

AAGAAAGTGA AACAGAAAAG GAAATTAAAA GGAAAGCTGA AGTTAAGAAA ACCAAAGCAG 240 GAAACACCAA AGAAGCAGTG GTTCACCTGA GAAAGAGCAC AAGAAACACA AGTAATATTC 300

CAGTGATTTT GGAACCTGAA ACTGAAGAAA GTGAAAATGA ATTTTATATC AAACAAAAGA 360

AAGCTAGACC TTCCGTCAAA GAAACTCTTC AGAAGTCTGG TGTTAGGAAA GAGTTTCCAA 420

TTACTGAGGC AGTAGGATCT GATAAGACAA ATAGGCATCC CTTAGAATGC TTACCTGGTT 480

TAATTCAGGA TAAGGAATGG AATGAGAAGG AGTTACAGAA ACTTCATTGT GCTTTTGCAT 540

CTCTTCCAAA GCACAAACCT GGTTTCTGGT CAGAGGTAGC TGCGGCTGTA GGTTCTCGAT 600

CTCCTGAAGA ATGCCAGAGG AAATACATGG AAAATCCCAG AGGAAAAGGA TCCCAGAAAC 660

ATGTCACTAA GAAGAAGCCA GCCAATTCCA AAGGCCAAAA TGGCAAGAGA GGTGATGCTG 720

ATCAGAAACA AACTATTAAG ATAACTGCCA AAGTGGGAAC TCTTAAAAGG AAGCAACAGA 780

TGAGGGAATT TCTGGAACAG TTGCCAAAAG ATGACCATGA TGATTTTTTC AGTACAACAC 840

CTTTACAGCA TCAAAGAATA CTGTTGCCAA GTTTCCAGGA CAGTGAAGAT GATGATGATA 900

TTCTGCCAAA TATGGACAAA AATCCAACAA CTCCATCATC AGTTATCTTT CCATTGGTAA 960

AAACTCCTCA ATGTCAGCAT GTCAGTCCTG GCATGCTAGG TTCTATAAAT AGGAATGACT 1020

GTGATAAATA TGTTTTTCGT ATGCAAAAAT ATCATAAAAG TAATGGTGGT ATTGTCTGGG 1080

GCAACATCAA GAAAAAATTA GTTGAAACTG ATTTCTCAAC TCCAACACCA AGAAGGAAAA 1140

CCCCATTTAA CACAGACTTA GGAGAAAACT CTGGTATTGG AAAACTTTTC ACTAATGCTG 1200

TGGAATCTTT AGATGAAGAA GAGAAAGATT ATTATTTTTC GAACTCTGAT TCTGCATAGT 1260

AAAATGAGAA AATATGATTC CTGGGATTTT TACCATAAAG CAGACAGTGT TTGTATTTTC 1320

AACTGGAGTA CATGTATTTT CTTTGTAAAG TAGCTTCCTA TGAAAATGTG GACTTTTTTG 1380

AAGGTTTCAT ATGTTTGTGT TCAAAGTAAA ATATCCTCAT TGCTGCAGCT TACTAAAAAT 1440

GTAAAGAAAA TTGTTTTTGC TCGTGTAGAT ATCTGTAAAT TTGTTTTTGC ATATTAAAAT 1500

A ATATAGAT AATTTTTTAA AAAAAAAAAA AAAAAAA 1537 (2) INFORMATION FOR SEQ ID NO : 4 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 397 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO : 4 :

Met Leu Thr Ser Asp Phe Lys Lys Asn Thr Arg Leu Leu Pro Lys Leu 1 5 10 15

Lys Lys lie Glu Asn Gin Val Ala Met Ser Phe Tyr Lys His Gin Ser 20 25 30

Ser Pro Asp Leu Ser Ser Glu Glu Ser Glu Thr Glu Lys Glu lie Lys 35 40 45

Arg Lys Ala Glu Val Lys Lys Thr Lys Ala Gly Asn Thr Lys Glu Ala 50 55 60

Val Val His Leu Arg Lys Ser Thr Arg Asn Thr Ser Asn lie Pro Val 65 70 75 80 lie Leu Glu Pro Glu Thr Glu Glu Ser Glu Asn Glu Phe Tyr lie Lys 85 90 95

Gin Lys Lys Ala Arg Pro Ser Val Lys Glu Thr Leu Gin Lys Ser Gly 100 105 110

Val Arg Lys Glu Phe Pro lie Thr Glu Ala Val Gly Ser Asp Lys Thr 115 120 125

Asn Arg His Pro Leu Glu Cys Leu Pro Gly Leu lie Gin Asp Lys Glu 130 135 140

Trp Asn Glu Lys Glu Leu Gin Lys Leu His Cys Ala Phe Ala Ser Leu 145 150 155 160

Pro Lys His Lys Pro Gly Phe Trp Ser Glu Val Ala Ala Ala Val Gly 165 170 175

Ser Arg Ser Pro Glu Glu Cys Gin Arg Lys Tyr Met Glu Asn Pro Arg 180 185 190

Gly Lys Gly Ser Gin Lys His Val Thr Lys Lys Lys Pro Ala Asn Ser 195 200 205

Lys Gly Gin Asn Gly Lys Arg Gly Asp Ala Asp Gin Lys Gin Thr lie 210 215 220

Lys lie Thr Ala Lys Val Gly Thr Leu Lys Arg Lys Gin Gin Met Arg 225 230 235 240

Glu Phe Leu Glu Gin Leu Pro Lys Asp Asp His Asp Asp Phe Phe Ser 245 250 255

Thr Thr Pro Leu Gin His Gin Arg lie Leu Leu Pro Ser Phe Gin Asp 260 265 270

Ser Glu Asp Asp Asp Asp lie Leu Pro Asn Met Asp Lys Asn Pro Thr 275 280 285

Thr Pro Ser Ser Val lie Phe Pro Leu Val Lys Thr Pro Gin Cys Gin 290 295 300

His Val Ser Pro Gly Met Leu Gly Ser lie Asn Arg Asn Asp Cys Asp 305 310 315 320

Lys Tyr Val Phe Arg Met Gin Lys Tyr His Lys Ser Asn Gly Gly lie 325 330 335

Val Trp Gly Asn lie Lys Lys Lys Leu Val Glu Thr Asp Phe Ser Thr 340 345 350

Pro Thr Pro Arg Arg Lys Thr Pro Phe Asn Thr Asp Leu Gly Glu Asn 355 360 365

Ser Gly lie Gly Lys Leu Phe Thr Asn Ala Val Glu Ser Leu Asp Glu 370 375 380

Glu Glu Lys Asp Tyr Tyr Phe Ser Asn Ser Asp Ser Ala 385 390 395

(2) INFORMATION FOR SEQ ID NO : 5 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 921 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 5 :

CGCAAGGACG CTGGGCCGGT GGGCGGGGGC CGGCAGGTGC TCCGCAGCCG TCTGTGCCAC 60

CCAGAGCCGG CGGGCCGCTA GGTCCCCGGA GACCCTGCTA TGGTGCGTGC GGGCGCCGTG 120

GGGGCTCATC TCCCCGCGTC CGGCTTGGAT ATCTTCGGGG ACCTGAAGAA GATGAACAAG 180

CGCCAGCTCT ATTACCAGGT TTTAAACTTC GCCATGATCG TGTCTTCTGC ACTCATGATA 240

TGGAAAGGCT TGATCGTGCT CACAGGCAGT GAGAGCCCCA TCGTGGTGGT GCTGAGTGGC 300

AGTATGGAGC CGGCCTTTCA CAGAGGAGAC CTCCTGTTCC TCACAAATTT CCGGGAAGAC 360

CCAATCAGAG CTGGTGAAAT AGTTGTTTTT AAAGTTGAAG GACGAGACAT TCCAATAGTT 420

CACAGAGTAA TCAAAGTTCA TGAAAAAGAT AATGGAGACA TCAAATTTCT GACTAAAGGA 480

GATAATAATG AAGTTGATGA TAGAGGCTTG TACAAAGAAG GCCAGAACTG GCTGGAAAAG 540 AAGGACGTGG TGGGAAGAGC AAGAGGGTTT TTACCATATG TTGGTATGGT CACCATAATA 600

ATGAATGACT ATCCAAAATT CAAGTATGCT CTTTTGGCTG TAATGGGTGC ATATGTGTTA 660

CTAAAACGTG AATCCTAAAA TGAGAAGCAG TTCCTGGGAC CAGATTGAAA TGAATTCTGT 720

TGAAAAAGAG AAAAACTAAT ATATTTGAGA TGTTCCATTT TCTGTATGAA AGGGAACAGT 780

GTGGAGATGT TTTTGTCTTG TCCAAATAAA AGATTCACCA GTAAAAAAAA AAAAAAAAAA 840

AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA 900

AAAAAAAAAA AAAAAAAAAA A 921 (2) INFORMATION FOR SEQ ID NO : 6 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 192 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 6 :

Met Val Arg Ala Gly Ala Val Gly Ala His Leu Pro Ala Ser Gly Leu 1 5 10 15

Asp lie Phe Gly Asp Leu Lys Lys Met Asn Lys Arg Gin Leu Tyr Tyr 20 25 30

Gin Val Leu Asn Phe Ala Met lie Val Ser Ser Ala Leu Met lie Trp 35 40 45

Lys Gly Leu lie Val Leu Thr Gly Ser Glu Ser Pro lie Val Val Val 50 55 60

Leu Ser Gly Ser Met Glu Pro Ala Phe His Arg Gly Asp Leu Leu Phe 65 70 75 80

Leu Thr Asn Phe Arg Glu Asp Pro lie Arg Ala Gly Glu lie Val Val 85 90 95

Phe Lys Val Glu Gly Arg Asp lie Pro lie Val His Arg Val lie Lys 100 105 110

Val His Glu Lys Asp Asn Gly Asp lie Lys Phe Leu Thr Lys Gly Asp 115 120 125

Asn Asn Glu Val Asp Asp Arg Gly Leu Tyr Lys Glu Gly Gin Asn Trp 130 135 140 Leu Glu Lys Lys Asp Val Val Gly Arg Ala Arg Gly Phe Leu Pro Tyr

145 150 155 160

Val Gly Met Val Thr lie lie Met Asn Asp Tyr Pro Lys Phe Lys Tyr

165 170 175

Ala Leu Leu Ala Val Met Gly Ala Tyr Val Leu Leu Lys Arg Glu Ser

180 185 190

(2) INFORMATION FOR SEQ ID NO : 7 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1505 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 7 :

GAACGGTGCT TTCAGGAACA ATAACAGTGG GGGGAGCCTG GGTCCTGGGG ARCCGCCCCC 60

GCCCCCCCCA GCACCGTAGC GAGACTCGCG CCCCCCGCCG GATCCCCACC TCCGCGCCCG 120

CCCCGCGGGC TGARCTGCCC GACCGGGGCC CGCCCCCGGC GCCCACCATG TACGCCTTTG 180

TKCGGTTCCT GGAGGACAAC GTCTGCTACG CGCTGCCCGT GTCGTGCGTG CGCGACTTCA 240

GCCCCCGCTC GCGGCTGGAT TTTGACAACC AGAAGGTGTA CGCCGTGTAC CGGGGCCCGG 300

AGGAATTGGG CGCCGGGCCC GAGAGCCCCC CGCGCGCCCC CCGCGACTGG GGCGCGCTGT 360

TGCTCCACAA GGCCCAGATC CTGGCGCTGG CAGAAGACAA ATCTGACCTT GAAAACAGTG 420

TGATGCAGAA GAAAATAAAA ATCCCCAAGC TTTCTCTTAA TCATGTAGAA GAAGATGGAG 480

AGGTTAAAGA TTATGGGGAA GAAGATTTAC AGCTTAGACA CATCAAGAGA CCTGAGGGGC 540

GGAAGCCGAG CGAAGTGGCG CACAAGAGCA TCGAGGCAGT GGTGGCTCGG CTAGAGAAGC 600

AGAACGGCCT GAGCCTGGGC CATAGCACGT GTCCGGAAGA GGTCTTCGTG GAGGCCTCGC 660

CAGGCACAGA GGACATGGAC AGTCTAGAAG ATGCTGTGGT GCCCCGGGCT CTGTATGAGG 720

AGCTGCTGCG CAACTACCAG CAGCAACAGG AAGAGATGCG CCACCTCCAG CAGGAGCTGG 780

AGCGGACTCG GAGGCAGCTG GTACAACAGG CCAAGAAGCT CAAGGAGTAC GGGGCACTTG 840

TGTCTGAAAT GAAGGAGCTC CGTGACCTTA ACCGGAGGCT CCAGGACGTG CTGCTCCTGA 900 GGCTTGGCAG CGGTCCCGCC ATTGATCTGG AAAAAGTAAA GTCAGAATGT CTCGAGCCCG 960

AGCCGGAGTT ACGGAGCACT TTCAGTGAGG AAGCAAATAC GTCGTCCTAT TACCCCGCTC 1020

CTGCGCCTGT CATGGACAAG TATATCCTAG ACAATGGCAA GGTCCATCTG GGAAGCGGGA 1080

TTTGGGTTGA TGAGGAGAAA TGGCACCAGC TACAAGTAAC CCAAGGAGAT TCCAAGTACA 1140

CGAAGAACTT GGCAGTTATG ATTTGGGGAA CAGATGTTCT GAAAAACAGA AGCGTCACAG 1200

GCGTCGCCAC AAAAAAAAAG AAAGATGCAG TCCCTAAACC ACCCCTCTCG CCTCACAAAC 1260

TAAGCATCGT CAGAGAGTGT TTGTATGACA GAATAGCACA AGAAACTGTG GATGAAACTG 1320

AAATTGCACA GAGACTCTCC AAAGTCAACA AGTACATCTG TGAAAAAATC ATGGATATCA 1380

ATAAATCCTG TAAAATGAAG AACGAAGGGA AGCAAAATAC AATTTGCAAT AAACTTTGGA 1440

TTTTTCATAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA 1500

AAAAA 1505 (2) INFORMATION FOR SEQ ID NO : 8 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 427 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 8 :

Met Tyr Ala Phe Val Arg Phe Leu Glu Asp Asn Val Cys Tyr Ala Leu 1 5 10 15

Pro Val Ser Cys Val Arg Asp Phe Ser Pro Arg Ser Arg Leu Asp Phe 20 25 30

Asp Asn Gin Lys Val Tyr Ala Val Tyr Arg Gly Pro Glu Glu Leu Gly 35 40 45

Ala Gly Pro Glu Ser Pro Pro Arg Ala Pro Arg Asp Trp Gly Ala Leu 50 55 60

Leu Leu His Lys Ala Gin lie Leu Ala Leu Ala Glu Asp Lys Ser Asp 65 70 75 80

Leu Glu Asn Ser Val Met Gin Lys Lys lie Lys lie Pro Lys Leu Ser 85 90 95 Leu Asn His Val Glu Glu Asp Gly Glu Val Lys Asp Tyr Gly Glu Glu 100 105 110

Asp Leu Gin Leu Arg His lie Lys Arg Pro Glu Gly Arg Lys Pro Ser 115 120 125

Glu Val Ala His Lys Ser lie Glu Ala Val Val Ala Arg Leu Glu Lys 130 135 140

Gin Asn Gly Leu Ser Leu Gly His Ser Thr Cys Pro Glu Glu Val Phe 145 150 155 160

Val Glu Ala Ser Pro Gly Thr Glu Asp Met Asp Ser Leu Glu Asp Ala 165 170 175

Val Val Pro Arg Ala Leu Tyr Glu Glu Leu Leu Arg Asn Tyr Gin Gin 180 185 190

Gin Gin Glu Glu Met Arg His Leu Gin Gin Glu Leu Glu Arg Thr Arg 195 200 205

Arg Gin Leu Val Gin Gin Ala Lys Lys Leu Lys Glu Tyr Gly Ala Leu 210 215 220

Val Ser Glu Met Lys Glu Leu Arg Asp Leu Asn Arg Arg Leu Gin Asp 225 230 235 240

Val Leu Leu Leu Arg Leu Gly Ser Gly Pro Ala lie Asp Leu Glu Lys 245 250 255

Val Lys Ser Glu Cys Leu Glu Pro Glu Pro Glu Leu Arg Ser Thr Phe 260 265 270

Ser Glu Glu Ala Asn Thr Ser Ser Tyr Tyr Pro Ala Pro Ala Pro Val 275 280 285

Met Asp Lys Tyr lie Leu Asp Asn Gly Lys Val His Leu Gly Ser Gly 290 295 300 lie Trp Val Asp Glu Glu Lys Trp His Gin Leu Gin Val Thr Gin Gly 305 310 315 320

Asp Ser Lys Tyr Thr Lys Asn Leu Ala Val Met lie Trp Gly Thr Asp 325 330 335

Val Leu Lys Asn Arg Ser Val Thr Gly Val Ala Thr Lys Lys Lys Lys 340 345 350

Asp Ala Val Pro Lys Pro Pro Leu Ser Pro His Lys Leu Ser lie Val 355 360 365

Arg Glu Cys Leu Tyr Asp Arg lie Ala Gin Glu Thr Val Asp Glu Thr 370 375 380

Glu lie Ala Gin Arg Leu Ser Lys Val Asn Lys Tyr lie Cys Glu Lys 385 390 395 400 lie Met Asp lie Asn Lys Ser Cys Lys Met Lys Asn Glu Gly Lys Gin 405 410 415

Asn Thr lie Cys Asn Lys Leu Trp lie Phe His 420 425

(2) INFORMATION FOR SEQ ID NO : 9 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2033 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 9:

GAAGCACGCT GAAACCCTGG GCGGCGGCAA GCTGTGCGAC CTCTTCTGCG GCCGGCCTGG 60

GCAGGTGTCT TCCTCGAGAG GCAGGCAGGG GATCCCGGAC CCTTATACAG GATGCTGTGT 120

TCTTTGCTCC TTTGTGAATG TCTGTTGCTG GTAGCTGGTT ATGCTCATGA TGATGACTGG 180

ATTGACCCCA CAGACATGCT TAACTATGAT GCTGCTTCAG GAACAATGAG AAAATCTCAG 240

GCAAAATATG GTATTTCAGG GGAAAAGGAT GTCAGTCCTG ACTTGTCATG TGCTGATGAA 300

A ATCAGAAT GTTATCACAA ACTTGATTCT TTAACTTATA AGATTGATGA RTGTKAAAAG 360

AAAAAGAGGG AAGACTATGA AAGTCAAAGC AATCCTGTTT TTAGGAGATA CTTAAATAAG 420

ATTTTAATTG AAGCTGGAAA GCTTGGACTT CCTGATGAAA ACAAAGGCGA TATGCATTAT 480

GATGCTGAGA TTATCCTTAA AAGAGAAACT TTGTTAGAAA TACAGAAGTT TCTCAATGGA 540

GAAGACTGGA AACCAGGTGC CTTGGATGAT GCACTAAGTG ATATTTTAAT TAATTTTAAG 600

TTTCATGATT TTGAAACATG GAAGTGGCGA TTCGAAGATT CCTTTGGAGT GGATCCATAT 660

AATGTGTTAA TGGTACTTCT TTGTCTGCTC TGCATCGTGG TTTTAGTGGC TACCGAGCTG 720

TGGACATATG TACGTTGGTA CACTCAGTTG AGACGTGTTT TAATCATCAG CTTTCTGTTC 780

AGTTTGGGAT GGAATTGGAT GTATTTATAT AAGCTAGCTT TTGCACAGCA TCAGGCTGAA 840

GTCGCCAAGA TGGAGCCATT AAACAATGTG TGTGCCAAAA AGATGGACTG GACTGGAAGT 900

ATCTGGGAAT GGTTTAGAAG TTCATGGACC TATAAGGATG ACCCATGCCA AAAATACTAT 960 GAGCTCTTAC TAGTCAACCC TATTTGGTTG GTCCCACCAA CAAAGGCACT TGCAGTTACA 1020

TTCACCACAT TTGTAACGGA GCCATTGAAG CATATTGGAA AAGGAACTGG GGAATTTATT 1080

AAAGCACTCA TGAAGGAAAT TCCAGCGCTG CTTCATCTTC CAGTGCTGAT AATTATGGCA 1140

TTAGCCATCC TGAGTTTCTG CTATGGTGCT GGAAAATCAG TTCATGTGCT GAGACATATA 1200

GGCGGTCCTT GAGAGAGAAC CTCCCCAGGC ACTTCGGCCA CGGGATAGAA GACGGCAGGA 1260

GGAAATTGAT TATAGACCTG ATGGTGGAGC AGGTGATGCC GATTTCCATT ATAGGGGCCA 1320

AATGGGCCCC ACTGAGCAAG GCCCTTATGC CAAAACGTAT GAGGGTAGAA GAGAGATTTT 1380

GAGAGAGAGA GATGTTGACT TGAGATTTCA GACTGGCAAC AAGAGCCCTG AAGTGCTCCG 1440

GGCATTTGAT GTACCAGACG CAGAGGCACG AGAGCATCCC ACGGTGGTAC CCAGTCATAA 1500

ATCACCTGTT TTGGATACAA AGCCCAAGGA GACAGGTGGA ATCCTGGGGG AAGGCACACC 1560

GAAAGAAAGC AGTACTGAAA GCAGCCAGTC GGCCAAGCCT GTCTCTGGCC AAGACACATC 1620

AGGGAATACA GAAGGTTCAC CCGCAGCGGA AAAGGCCCAG CTCAAGTCTG AAGCCGCAGG 1680

CAGCCCAGAC CAAGGCAGCA CATACAGCCC CGCAAGAGGT GTGGCTGGAC CACGTGGACA 1740

GGATCCGGTC AGCAGCCCCT GTGGCTAGAG GAACACCAGC ACAAASGACA GCCTCAAGTC 1800

TCCTTCGAGC TTTATATCCA TTTGGGGATG AAGTYTAYTT TGACAGCTAG CAAGGCGACA 1860

TGCAACTGTT GTTGAATGAT GACAGCAATT CAGGAAAGAY TTAAATATGA AAGCAAATTG 1920

AACACATCGG GTGTTTGTTA TCAGAAAAGA GATGAGATGA GATAAGACTT GTTTATTGAC 1980

TAGCCAATAT GTCATTAAAA TTAAGGTTTA TATTGTGAAA AAAAAAAAAA AAA 2033 (2) INFORMATION FOR SEQ ID NO: 10:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 366 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10:

Met Leu Cys Ser Leu Leu Leu Cys Glu Cys Leu Leu Leu Val Ala Gly 1 5 10 15

Tyr Ala His Asp Asp Asp Trp lie Asp Pro Thr Asp Met Leu Asn Tyr 20 25 30

Asp Ala Ala Ser Gly Thr Met Arg Lys Ser Gin Ala Lys Tyr Gly lie 35 40 45

Ser Gly Glu Lys Asp Val Ser Pro Asp Leu Ser Cys Ala Asp Glu lie 50 55 60

Ser Glu Cys Tyr His Lys Leu Asp Ser Leu Thr Tyr Lys lie Asp Glu 65 70 75 80

Cys Xaa Lys Lys Lys Arg Glu Asp Tyr Glu Ser Gin Ser Asn Pro Val 85 90 95

Phe Arg Arg Tyr Leu Asn Lys lie Leu lie Glu Ala Gly Lys Leu Gly 100 105 110

Leu Pro Asp Glu Asn Lys Gly Asp Met His Tyr Asp Ala Glu lie lie 115 120 125

Leu Lys Arg Glu Thr Leu Leu Glu lie Gin Lys Phe Leu Asn Gly Glu 130 135 140

Asp Trp Lys Pro Gly Ala Leu Asp Asp Ala Leu Ser Asp lie Leu lie 145 150 155 160

Asn Phe Lys Phe His Asp Phe Glu Thr Trp Lys Trp Arg Phe Glu Asp 165 170 175

Ser Phe Gly Val Asp Pro Tyr Asn Val Leu Met Val Leu Leu Cys Leu 180 185 190

Leu Cys lie Val Val Leu Val Ala Thr Glu Leu Trp Thr Tyr Val Arg 195 200 205

Trp Tyr Thr Gin Leu Arg Arg Val Leu lie lie Ser Phe Leu Phe Ser 210 215 220

Leu Gly Trp Asn Trp Met Tyr Leu Tyr Lys Leu Ala Phe Ala Gin His 225 230 235 240

Gin Ala Glu Val Ala Lys Met Glu Pro Leu Asn Asn Val Cys Ala Lys 245 250 255

Lys Met Asp Trp Thr Gly Ser lie Trp Glu Trp Phe Arg Ser Ser Trp 260 265 270

Thr Tyr Lys Asp Asp Pro Cys Gin Lys Tyr Tyr Glu Leu Leu Leu Val 275 280 285

Asn Pro lie Trp Leu Val Pro Pro Thr Lys Ala Leu Ala Val Thr Phe 290 295 300

Thr Thr Phe Val Thr Glu Pro Leu Lys His lie Gly Lys Gly Thr Gly 305 310 315 320 Glu Phe lie Lys Ala Leu Met Lys Glu lie Pro Ala Leu Leu His Leu 325 330 335

Pro Val Leu lie lie Met Ala Leu Ala lie Leu Ser Phe Cys Tyr Gly 340 345 350

Ala Gly Lys Ser Val His Val Leu Arg His lie Gly Gly Pro 355 360 365

(2) INFORMATION FOR SEQ ID NO: 11:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2214 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11:

GTGAGCTGCA AATCTTGGAG CAAAAACCAG AGACATTGCC AGAGCAAACA AGAACAGAAA 60

TACAAATGGA GAACTGGTCA AAAGTTTGCT ACTTCGCATA TCCCTACTTG ACCTTTTTTG 120

GAGCAAATGC CTTTGTGAAG TCTTCCTAGA GTCTTCCAGA CATAACCCAC AGTTATCTTG 180

AACAAGAAAC TACGGGGATA AATAAAAGTA CGCAGCCAGA TGAGCAACTG ACTATGAATT 240

CTGAGAAAAG TATGCATCGG AAATCCACTG AATTAGTTAA TGAAATAACA TGTGAGAACA 300

CAGAATGGCC AGGGCAGAGA TCAACGAATT TTCAGATCAT CAGTTCTTAT CCAGATGATG 360

AGTCTGTTTA CTGCACTACT GAAAAATACA ACGTTATGGA ACATAGACAT AATGATATGC 420

ATTATGAATG TWTGACTCCT TGTCAAGTTA CTTCAGACTC AGATAAAGAG AAGACAATAG 480

CATTTCTTCT AAAAGAATTG GATATTCTCA GAACAAGCAA TAAAAAGCTT CAGCAGAAAT 540

TGGCTAAAGA AGATAAAGAA CAGAGAAAAC TAAAGTTTAA GCTGGAACTC CAAGAGAAAG 600

AAACAGAAGC TAAAATTGCT GAAAAGACAG CAGCTCTGGT TGAAGAAGTG TATTTTGCGC 660

AGAAGGAACG TGATGAAGCT GTTATGTCTA GACTGCAATT AGCCATTGAG GAGAGAGATG 720

AAGCAATTGC ACGAGCCAAG CATATGGAAA TGTCTCTAAA AGTGCTAGAA AATATTAACC 780

CTGAAGAAAA TGACATGACA TTACAGGAAT TACTGAACAG AATAAACAAT GCAGACACAG 840

GGATAGCTAT TCAGAAGAAT GGAGCTATAA TTGTGGATAG AATCTACAAG ACCAAGGAAT 900

GTAAAATGAG AATAACTGCA GAAGAAATGA GTGCACTAAT AGAAGAACGG GATGCTGCCT 960 TGTCTAAGTG CAAACGGTTA GAGCAGGAGC TTCATCATGT GAAAGAGCAG AACCAGACTT 1020

CAGCAAACAA CATGAGACAT CTGACTGCTG AAAACAATCA AGAACGTGCT CTGAAGGCAA 1080

AGTTGTTATC TATGCAACAA GCCAGAGAAA CTGCAGTTCA ACAGTACAAA AAACTGGAAG 1140

AGGAAATCCA GACCCTTCGA GTTTACTACA GTTTACACAA ATCTTTATCT CAAGAAGAAA 1200

ATCTGAAGGA TCAGTTTAAC TATACCCTTA GTACATATGA AGAAGCTTTA AAAAACAGAG 1260

AGAACATTGT TTCCATCACT CAACAACAAA ATGAGGAACT GGCTACTCAA CTGCAACAAG 1320

CTCTGACAGA GCGAGCAAAT ATGGAATTAC AACTTCAACA TGCCAGAGAG GCCTCCCAAG 1380

TGGCCAATGA AAAAGTTCAA AAGTTGGAAA GGCTGGTGGA TGTACTGAGG AAGAAGGTTG 1440

GAACCGGGAC CATGAGGACA GTGATCTGAT TGAAAAAAAC GACAGTCTGG GGAAGCGATC 1500

ACATCTGGTG ACCAGGCTGC TTCATTCAAC ACTGTGTAAA CACCAAAGCC TTAACTTAGC 1560

AAACAGTTGT TAGAAGTGGG ACACTCCAAC CACATTCCAA GCTGAGATAA AATCAAATCA 1620

CAAATGTTTA ACCACTTTGC TGCTGACTTG AGTTATTTAT CCAAATATAT TAACTATAGA 1680

CTTTTACCAA TGGGTAGCTA TAAGGTTACA GCTTATTTTG TAACTATTTT ATATCTCAAT 1740

ATCTTTAATA TAAATCTTTT TACTGAGAGA TCATTATAGA AACATGTTAA AGTTGGTTAG 1800

GATCATATCT TCACATATGG CCCTTTCTGA ATCAAAGTGC GGCAAAGTAA ATATTGTCTA 1860

AGCTTTAATC CACTGTGTTA GGTCAAAACT TCAAATACAT GCATTTTTCA ATATAGGGTA 1920

CATTTCTTAA CTGATGAGAG AGGCTTAGAC ATGAGTGTGT AGTCTTCCTT CAATGCGTGT 1980

ATGTAATCTT TGTTAGTATA AAAGATATTA AATATAGGTG CCAAGAATTA AATGTATAAT 2040

TTGTTTAATA AGAGATGGAT ATATTAAAAT TACATTCATC AAGGCATGAT TTTTGTTTCA 2100

CTACAAATAA TGCAAACTGT TTTCAATAAA AAGAGGAGAC TGTTAATGTK KTAAAAAAAA 2160

AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAA 2214 (2) INFORMATION FOR SEQ ID NO: 12:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 411 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:

Met Asn Ser Glu Lys Ser Met His Arg Lys Ser Thr Glu Leu Val Asn 1 5 10 15

Glu lie Thr Cys Glu Asn Thr Glu Trp Pro Gly Gin Arg Ser Thr Asn 20 25 30

Phe Gin lie lie Ser Ser Tyr Pro Asp Asp Glu Ser Val Tyr Cys Thr 35 40 45

Thr Glu Lys Tyr Asn Val Met Glu His Arg His Asn Asp Met His Tyr 50 55 60

Glu Cys Xaa Thr Pro Cys Gin Val Thr Ser Asp Ser Asp Lys Glu Lys 65 70 75 80

Thr lie Ala Phe Leu Leu Lys Glu Leu Asp lie Leu Arg Thr Ser Asn 85 90 95

Lys Lys Leu Gin Gin Lys Leu Ala Lys Glu Asp Lys Glu Gin Arg Lys 100 105 110

Leu Lys Phe Lys Leu Glu Leu Gin Glu Lys Glu Thr Glu Ala Lys lie 115 120 125

Ala Glu Lys Thr Ala Ala Leu Val Glu Glu Val Tyr Phe Ala Gin Lys 130 135 140

Glu Arg Asp Glu Ala Val Met Ser Arg Leu Gin Leu Ala lie Glu Glu 145 150 155 160

Arg Asp Glu Ala lie Ala Arg Ala Lys His Met Glu Met Ser Leu Lys 165 170 175

Val Leu Glu Asn lie Asn Pro Glu Glu Asn Asp Met Thr Leu Gin Glu 180 185 190

Leu Leu Asn Arg lie Asn Asn Ala Asp Thr Gly lie Ala lie Gin Lys 195 200 205

Asn Gly Ala lie lie Val Asp Arg lie Tyr Lys Thr Lys Glu Cys Lys 210 215 220

Met Arg lie Thr Ala Glu Glu Met Ser Ala Leu lie Glu Glu Arg Asp 225 230 235 240

Ala Ala Leu Ser Lys Cys Lys Arg Leu Glu Gin Glu Leu His His Val 245 250 255

Lys Glu Gin Asn Gin Thr Ser Ala Asn Asn Met Arg His Leu Thr Ala 260 265 270

Glu Asn Asn Gin Glu Arg Ala Leu Lys Ala Lys Leu Leu Ser Met Gin 275 280 285 Gin Ala Arg Glu Thr Ala Val Gin Gin Tyr Lys Lys Leu Glu Glu Glu 290 295 300 lie Gin Thr Leu Arg Val Tyr Tyr Ser Leu His Lys Ser Leu Ser Gin 305 310 315 320

Glu Glu Asn Leu Lys Asp Gin Phe Asn Tyr Thr Leu Ser Thr Tyr Glu 325 330 335

Glu Ala Leu Lys Asn Arg Glu Asn lie Val Ser lie Thr Gin Gin Gin 340 345 350

Asn Glu Glu Leu Ala Thr Gin Leu Gin Gin Ala Leu Thr Glu Arg Ala 355 360 365

Asn Met Glu Leu Gin Leu Gin His Ala Arg Glu Ala Ser Gin Val Ala 370 375 380

Asn Glu Lys Val Gin Lys Leu Glu Arg Leu Val Asp Val Leu Arg Lys 385 390 395 400

Lys Val Gly Thr Gly Thr Met Arg Thr Val lie 405 410

(2) INFORMATION FOR SEQ ID NO: 13:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1227 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:

CCCAGCTACT CGCAAGGCTG AGGCAGGAGA ATCGCTTGAA CCTGGGAGGC GGAGGTTGCA 60

GTGAGCCAAG ATTGCGCCAT TGCACTCCAG CCTGGGCAAC AAGAGAGAAA CTCTGTCTCA 120

AAAAAAAAAA AAAAAAAAAA AAAAAAAAAG TCTGTTCTGT GGCCTCCTTC CCCCGGGCCC 180

TCCATAACCT CAGGCTAAAC CATGAACACA TCTGTATATC CTTGGCTCCA CTAAATGGCC 240

CTCTTGTGGC CCTCCAAACC ATGCTGCCTA TACACAGTGC TCTGTGGTAT GAAGCAGAGA 300

TTTTTCCTTT GCTCATTTTT AACTTTTGCC ATTGTGGTAG GTTGCAGCCA TCTTTCTAGA 360

TGTAGTGTAA CTGATGTAGT GTAACTGAAT GATGGTCACC TGCCATTGAG TGAGAGGTTG 420

AGTGCCATCC TTACTTGTCC ACTGTGGGGA AAGATAAAGC AAGCCTGGGT CTTTGCTAAG 480 GAAATAATTG TACCTGAAAG AAGCAAAAGC AGGGCTATCC AAAGGTGCTA CTGTGAGCAT 540

CCATGAAGAA GGAACAGTTA CCATCTCTCA GCTCCTACAC ACCTGGTCTC TGATGGGCCC 600

CTGACTGCTG CACTCTCTCA GTGGGGCAAG GTTGGAGATG GGCTAAGGAT CTGTATAGTA 660

GTAGCAGGTC TCAGTGGCTT TCTATTCGCT CAGGTTCCCT TGTTCTGTGG TCCTGGTCGG 720

GAGCAGGCCC TGGAGCTGTG GATGAAGCCA CTTGAGTGGC AGTAGCCGGC CCCCCTTTCC 780

TGTGCCTTCC CTTTGCACAA AGACACGTTC CTTCTGCGCT TGGCAGGTGC TAACTTCATG 840

GAAGATGGTA ATAGGAACAG ATACTGCTCT CTGCCCCTGC TTCTTGGATT CTTCAGCTCC 900

TACCGCTGAT TCGGATTATC CCCTTTTGAC CCTCTTGGGG ATCCAGACTG CTGCCAGGAG 960

GCTGAGTAGA TACCTCAGTT ATCTGTATGT TATTTCTGAG AGGCTTTGGC GGATCCTCTT 1020

CACTTGGCAT CCTGTGGCAT CCCTAGGGTC TGGTGGTAGC AGTAATAGCA ACAATGATAT 1080

CTTACATCTG GACAACTCAG AATAGTGGTA ACTTTTCCTC CTGACCTTTG CAAAATGACA 1140

TCATTTTCCC TTGTACATTT GATACCCTTG CTAGTAGAGA AGGGGAAATG CTTAAAGATG 1200

AAAATTCAAG TGAAAAAAAA AAAAAAA 1227 (2) INFORMATION FOR SEQ ID NO: 14:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 86 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 14:

Met Val lie Gly Thr Asp Thr Ala Leu Cys Pro Cys Phe Leu Asp Ser 1 5 10 15

Ser Ala Pro Thr Ala Asp Ser Asp Tyr Pro Leu Leu Thr Leu Leu Gly 20 25 30 lie Gin Thr Ala Ala Arg Arg Leu Ser Arg Tyr Leu Ser Tyr Leu Tyr 35 40 45

Val lie Ser Glu Arg Leu Trp Arg lie Leu Phe Thr Trp His Pro Val 50 55 60

Ala Ser Leu Gly Ser Gly Gly Ser Ser Asn Ser Asn Asn Asp lie Leu 65 70 75 80 His Leu Asp Asn Ser Glu 85

(2) INFORMATION FOR SEQ ID NO: 15:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1232 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15:

CAGGCTTAGA GACGGCCTCG TGGGTGCCCA GTTCTGGTCA GCCTCCGTCT CATGCCAGTC 60

CCAGGACCAG ACTGCCGTGC GCCTCGCCCT GGAGCAGATT GACCTCATTC ACCGCATGTG 120

TGCCTCCTAC TCTGAACTCG AGCTTGTGAC CTCAGCTGAA GGTCTGAACA GCTCTCAAAA 180

GCTGGCCTGC CTCATTGGCG TGGAGGGTGG TCACTCACTG GACAGCAGCC TCTCTGTGCT 240

GCGCAGTTTC TATGTGCTGG GGGTGCGCTA CCTGACACTT ACCTTCACCT GCAGTACACC 300

ATGGGCAGAG AGTTCCACCA AGTTCAGACA CCACATGTAC ACCAACGTCA GCGGATTGAC 360

AAGCTTTGGT GAGAAAGTAG TAGAGGAGTT GAACCGCCTG GGCATGATGA TAGATTTGTC 420

CTATGCATCG GACACCTTGA TAAGAAGGGT CCTGGAAGTG TCTCAGGCTC CTGTGATCTT 480

CTCCCACTCA GCTGCCAGAG CTGTGTGTGA CAATTTGTTG AATGTTCCCG ATGATATCCT 540

GCAGCTTCTG AAGAAGAACG GTGGCATCGT GATGGTGACA CTGTCCATGG GGGTGCTGCA 600

GTGCAACCTG CTTGCTAACG TGTCCACTGT GGCAGATCAC TTTGACCACA TCAGGGCAGT 660

CATTGGATCT GAGTTCATCG GGATTGGTGG AAATTATGAC GGGACTGGCC GGTTCCCTCA 720

GGGGCTGGAG GATGTGTCCA CATACCCAGT CCTGATAGAG GAGTTGCTGA GTCGTAGCTG 780

GAGCGAGGAA GAGCTTCAAG GTGTCCTTCG TGGAAACCTG CTGCGGGTCT TCAGACAAGT 840

GGAAAAGGTG AGAGAGGAGA GCAGGGCGCA GAGCCCCGTG GAGGCTGAGT TTCCATATGG 900

GCAACTGAGC ACATCCTGCC ACTCCCACCT CGTGCCTCAG AATGGACACC AGGCTAATCA 960

TCTGGAGGTG ACCAAGCAGC CAACCAATCG GGTCCCCTGG AGGTCCTCAA ATGCCTCCCC 1020

ATACCTTGTT TCAGGCCTTG TGGGTGGTGC CACCATCCCA ACCTTCACCC AGTGGTTTTG 1080

CTGAAACAGT CGGTCCCCGC AGAGGTCACT GTGGCAAAGC CTCACAAAGC CCCCTTTCCT 1140 AGTTCATTCA CAAGCATATG CTGAGAATAA ACATGTTACA CATGGGAAAA AAAAAAAAAA 1200 AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AA 1232

(2) INFORMATION FOR SEQ ID NO: 16:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 322 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 16:

Met Cys Ala Ser Tyr Ser Glu Leu Glu Leu Val Thr Ser Ala Glu Gly 1 5 10 15

Leu Asn Ser Ser Gin Lys Leu Ala Cys Leu lie Gly Val Glu Gly Gly 20 25 30

His Ser Leu Asp Ser Ser Leu Ser Val Leu Arg Ser Phe Tyr Val Leu 35 40 45

Gly Val Arg Tyr Leu Thr Leu Thr Phe Thr Cys Ser Thr Pro Trp Ala 50 55 60

Glu Ser Ser Thr Lys Phe Arg His His Met Tyr Thr Asn Val Ser Gly 65 70 75 80

Leu Thr Ser Phe Gly Glu Lys Val Val Glu Glu Leu Asn Arg Leu Gly 85 90 95

Met Met lie Asp Leu Ser Tyr Ala Ser Asp Thr Leu lie Arg Arg Val 100 105 110

Leu Glu Val Ser Gin Ala Pro Val lie Phe Ser His Ser Ala Ala Arg 115 120 125

Ala Val Cys Asp Asn Leu Leu Asn Val Pro Asp Asp lie Leu Gin Leu 130 135 140

Leu Lys Lys Asn Gly Gly lie Val Met Val Thr Leu Ser Met Gly Val 145 150 155 160

Leu Gin Cys Asn Leu Leu Ala Asn Val Ser Thr Val Ala Asp His Phe 165 170 175

Asp His lie Arg Ala Val lie Gly Ser Glu Phe lie Gly lie Gly Gly 180 185 190 Asn Tyr Asp Gly Thr Gly Arg Phe Pro Gin Gly Leu Glu Asp Val Ser 195 200 205

Thr Tyr Pro Val Leu lie Glu Glu Leu Leu Ser Arg Ser Trp Ser Glu 210 215 220

Glu Glu Leu Gin Gly Val Leu Arg Gly Asn Leu Leu Arg Val Phe Arg 225 230 235 240

Gin Val Glu Lys Val Arg Glu Glu Ser Arg Ala Gin Ser Pro Val Glu 245 250 255

Ala Glu Phe Pro Tyr Gly Gin Leu Ser Thr Ser Cys His Ser His Leu 260 265 270

Val Pro Gin Asn Gly His Gin Ala Asn His Leu Glu Val Thr Lys Gin 275 280 285

Pro Thr Asn Arg Val Pro Trp Arg Ser Ser Asn Ala Ser Pro Tyr Leu 290 295 300

Val Ser Gly Leu Val Gly Gly Ala Thr lie Pro Thr Phe Thr Gin Trp 305 310 315 320

Phe Cys

(2) INFORMATION FOR SEQ ID NO: 17:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 3320 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 17:

CAGAGTTCTC CCTAAAGCTG CTGTCTTACT CTGTCAACGT GATAGTGGAC ATCCACGCAG 60

TGCAGCTCCT CTGGCACCAG CTTCGAGTCT CAGTGCTGGT TCTGCGGGAG CGCATTCTGC 120

AAGGTCTGCA GGACGCCAAT GGCAACTACA CTAGGCAGAC GGACATTCTG CAAGCTTTCT 180

CTGAAGAGAC AAAAGAGGGC CGGCTTGATT CTCTAACAGA AGTGGATGAC TCAGGACAAT 240

TAACCATCAA ATGTTCTCAA AATTACTTGT CTCTGGATTG TGGCATTACT GCATTCGAAC 300

TGTCTGACTA CAGTCCAAGT GAGGATTTGC TCAGTGGGCT AGGTGACATG ACCTCTAGCC 360

AAGTCAAAAC CAAACCCTTT GACTCTTGGA GCTACAGTGA GATGGAAAAG GAGTTTCCTG 420 AGCTTATCCG AAGTGTTGGT TTACTTACGG TAGCTGCTGA CTCTATCTCT ACCAATGGCA 480

GTGAAGCAGT TACTGAGGAG GTATCTCAAG TATCTCTCTC AGTAGACGAC AAAGGTGGAT 540

GTGAGGAAGA CAATGCTTCT GCAGTCGAAG AGCAACCAGG CTTAACACTG GGGGTGTCAT 600

CATCTTCAGG AGAAGCTCTG ACAAATGCTG TTCAACCCTC CTCTGAGACT GTGCAGCAAG 660

AATCCAGTTC CTCCTCCCAT CATGATGCAA AGAATCAGCA GCCTGTTCCT TGTGAAAATG 720

CAACCCCCAA ACGAACCATC AGAGATTGCT TTAATTATAA CGAGGACTCT CCCACACAGC 780

CTACATTGCC AAAAAGAGGA CTTTTTCTTA AAGAGGAAAC TTTTAAGAAT GATCTGAAAG 840

GCAATGGTGG AAAGAGGCAA ATGGTTGATC TAAAGCCTGA GATGAGCAGA AGCACCCCTT 900

CGCTAGTAGA TCCTCCTGAC AGATCCAAAC TTTGCCTGGT ATTGCAGTCT TCTTACCCCA 960

ACAGCCCTTC TGCTGCCAGC CAGTCTTATG AGTGTTTACA CAAGGTGGGG AATGGGAACC 1020

TTGAAAACAC AGTCAAATTT CACATTAAAG AAATTTCTTC CAGCCTGGGA AGGCTTAACG 1080

ACTGCTATAA AGAGAAATCT CGACTTAAAA AGCCACACAA GACCTCAGAA GAGGTGCCTC 1140

CATGCCGAAC ACCTAAACGG GGGACTGGTT CAGGCAAACA AGCTAAAAAT ACAAAGAGCT 1200

CAGCAGTGCC AAATGGAGAG CTTTCTTATA CTTCCAAGGC CATAGAGGGG CCACAAACAA 1260

ATTCTGCTTC CACATCCTCA CTTGAGCCTT GTAATCAGAG AAGTTGGAAT GCCAAATTGC 1320

AATTGCAGTC AGAAACATCC AGTTCACCAG CTTTTACTCA GAGCAGTGAA TCCTCTGTTG 1380

GCTCAGACAA CATCATGTCT CCGGTGCCAC TTCTTTCAAA ACACAAAAGC AAAAAAGGTC 1440

AAGCCTCCTC TCCAAGTCAC GTCACTAGGA ATGGTGAGGT TGTGGAGGCC TGGTATGGCT 1500

CTGATGAATA CCTAGCACTG CCCTCTCACC TTAAGCAGAC AGAAGTATTG GCTTTGAAGT 1560

TGGAAAACCT AACAAAGCTT CTGCCTCAGA AACCCAGAGG AGAAACCATC CAGAATATTG 1620

ATGACTGGGA ACTGTCTGAA ATGAATTCAG ATTCTGAAAT CTATCCAACC TATCATGTCA 1680

AAAAGAAGCA TACAAGGCTA GGCAGGGTGT CTCCAAGCTC ATCTAGTGAC ATAGCCTCTT 1740

CACTAGGGGA GAGCATTGAA TCTGGGCCCC TGAGTGACAT TCTTTCTGAT GAGGAGTCCA 1800

GTATGCCTCT CGCTGGCATG AAAAAGTATG CTGATGAGAA GTCAGAAAGA GCTTCATCCT 1860

CTGAGAAAAA TGAGAGCCAT TCTGCCACTA AATCAGCTTT AATTCAGAAA CTGATGCAAG 1920

ATATTCAGCA CCAAGACAAC TATGAAGCCA TATGGGAAAA AATAGAGGGG TTTGTAAACA 1980

AACTGGATGA ATTCATTCAA TGGTTAAATG AAGCCATGGA AACTACAGAA AATTGGACTC 2040

CCCCTAAAGC AGAGATGGAT GACCTTAAAC TGTATCTGGA GACACACTTG AGTTTTAAGT 2100 TGAATGTAGA CAGTCATTGT GCTCTCAAGG AAGCTGTGGA GGAGGAAGGA CACCAACTTC 2160

TTGAGCTTAT TGCATCTCAC AAAGCAGGAC TGAAGGACAT GCTGCGGATG ATTGCAAGTC 2220

AATGGAAGGA GCTGCAGAGG CAAATCAAAC GGCAGCACAG CTGGATTCTC AGGGCTCTGG 2280

ATACCATCAA AGCCGAGATA CTGGCTACTG ATGTGTCTGT GGAGGATGAG GAAGGGACTG 2340

GAAGCCCCAA GGCTGAGGTT CAACTATGCT ACCTGGAAGC ACAAAGAGAT GCTGTTGAGC 2400

AGATGTCCCT CAAGCTGTAC AGCGAGCAGT ATACCAGCAG CAGCAAGCGA AAGGAAGAGT 2460

TTGCTGATAT GTCAAAAGTT CATTCAGTGG GAAGCAATGG GCTTCTGGAC TTTGATTCAG 2520

AATATCAGGA GCTCTGGGAT TGGCTGATTG ACATGGAGTC CCTTGTGATG GACAGCCACG 2580

ACCTGATGAT GTCAGAGGAG CAGCAGCAGC ATCTTTACAA GGTTAGAGCT ACCCTTCCTG 2640

CCTTTACCTT GCTGTGGAAG ATCTGATTAG CCTGACAAGT CTCTCTCTCT CTCTCAGGAT 2700

ATCTTTTCGT TCATTGTATG TATCATGGTG TCTATTAGAA TTCCCTTCCC TGTCCCCAAA 2760

CTCATTTCCA TCCCTTGTGT GCTGACAGGC TGCACCGACA TCATAATTGC AAGAAAGTTC 2820

CCTTTATCAC ATTCTATTTG CTGTAATTCT ATAGAAGGAC AAAGATTTAT CTTCAAGATG 2880

AATAGCAATA AATGAAACTG CATTCATAAA TAGACTGAAA CAAAATGAAG GATAAATGGA 2940

CTAGTAATGT TTAACAAAAC GTGGTACGAT TGGAGCACTC AGGTGGATTC TATCTACCCC 3000

TCAGATCTTC CCAGGGAAAA AAGAAATTTG CCTTTGGTTT GGCTTGCTGC TGCCAAAGTA 3060

CAGGCCTCAT CCAATTTAAT GTTTATTAGT GATACATAGA GCAGACGTTG TCATATTAAG 3120

TCAATACATT TCCATGTGAT TTAAGAAAAC TGGCTTGAAA CTCATGTATT GATTGTAGGC 3180

AATCCATCTA ATAACTATCT ATATTGTTTT CCATGTTTGC TAATTCACCC TTAACCTACT 3240

GTATTTTACC AATTCTACTT AAATAGTCTT AGCAGTTCTT ATAAAGCAAT CATGTGATTT 3300

TGCCTAAAAA AAAAAAAAAA 3320 (2) INFORMATION FOR SEQ ID NO: 18:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 772 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 18:

Met Thr Ser Ser Gin Val Lys Thr Lys Pro Phe Asp Ser Trp Ser Tyr 1 5 10 15

Ser Glu Met Glu Lys Glu Phe Pro Glu Leu He Arg Ser Val Gly Leu 20 25 30

Leu Thr Val Ala Ala Asp Ser He Ser Thr Asn Gly Ser Glu Ala Val 35 40 45

Thr Glu Glu Val Ser Gin Val Ser Leu Ser Val Asp Asp Lys Gly Gly 50 55 60

Cys Glu Glu Asp Asn Ala Ser Ala Val Glu Glu Gin Pro Gly Leu Thr 65 70 75 80

Leu Gly Val Ser Ser Ser Ser Gly Glu Ala Leu Thr Asn Ala Val Gin 85 90 95

Pro Ser Ser Glu Thr Val Gin Gin Glu Ser Ser Ser Ser Ser His His 100 105 110

Asp Ala Lys Asn Gin Gin Pro Val Pro Cys Glu Asn Ala Thr Pro Lys 115 120 125

Arg Thr He Arg Asp Cys Phe Asn Tyr Asn Glu Asp Ser Pro Thr Gin 130 135 140

Pro Thr Leu Pro Lys Arg Gly Leu Phe Leu Lys Glu Glu Thr Phe Lys 145 150 155 160

Asn Asp Leu Lys Gly Asn Gly Gly Lys Arg Gin Met Val Asp Leu Lys 165 170 175

Pro Glu Met Ser Arg Ser Thr Pro Ser Leu Val Asp Pro Pro Asp Arg 180 185 190

Ser Lys Leu Cys Leu Val Leu Gin Ser Ser Tyr Pro Asn Ser Pro Ser 195 200 205

Ala Ala Ser Gin Ser Tyr Glu Cys Leu His Lys Val Gly Asn Gly Asn 210 215 220

Leu Glu Asn Thr Val Lys Phe His He Lys Glu He Ser Ser Ser Leu 225 230 235 240

Gly Arg Leu Asn Asp Cys Tyr Lys Glu Lys Ser Arg Leu Lys Lys Pro 245 250 255

His Lys Thr Ser Glu Glu Val Pro Pro Cys Arg Thr Pro Lys Arg Gly 260 265 270

Thr Gly Ser Gly Lys Gin Ala Lys Asn Thr Lys Ser Ser Ala Val Pro 275 280 285 Asn Gly Glu Leu Ser Tyr Thr Ser Lys Ala He Glu Gly Pro Gin Thr 290 295 300

Asn Ser Ala Ser Thr Ser Ser Leu Glu Pro Cys Asn Gin Arg Ser Trp 305 310 315 320

Asn Ala Lys Leu Gin Leu Gin Ser Glu Thr Ser Ser Ser Pro Ala Phe 325 330 335

Thr Gin Ser Ser Glu Ser Ser Val Gly Ser Asp Asn He Met Ser Pro 340 345 350

Val Pro Leu Leu Ser Lys His Lys Ser Lys Lys Gly Gin Ala Ser Ser 355 360 365

Pro Ser His Val Thr Arg Asn Gly Glu Val Val Glu Ala Trp Tyr Gly 370 375 380

Ser Asp Glu Tyr Leu Ala Leu Pro Ser His Leu Lys Gin Thr Glu Val 385 390 395 400

Leu Ala Leu Lys Leu Glu Asn Leu Thr Lys Leu Leu Pro Gin Lys Pro 405 410 415

Arg Gly Glu Thr He Gin Asn He Asp Asp Trp Glu Leu Ser Glu Met 420 425 430

Asn Ser Asp Ser Glu He Tyr Pro Thr Tyr His Val Lys Lys Lys His 435 440 445

Thr Arg Leu Gly Arg Val Ser Pro Ser Ser Ser Ser Asp He Ala Ser 450 455 460

Ser Leu Gly Glu Ser He Glu Ser Gly Pro Leu Ser Asp He Leu Ser 465 470 475 480

Asp Glu Glu Ser Ser Met Pro Leu Ala Gly Met Lys Lys Tyr Ala Asp 485 490 495

Glu Lys Ser Glu Arg Ala Ser Ser Ser Glu Lys Asn Glu Ser His Ser 500 505 510

Ala Thr Lys Ser Ala Leu He Gin Lys Leu Met Gin Asp He Gin His 515 520 525

Gin Asp Asn Tyr Glu Ala He Trp Glu Lys He Glu Gly Phe Val Asn 530 535 540

Lys Leu Asp Glu Phe He Gin Trp Leu Asn Glu Ala Met Glu Thr Thr 545 550 555 560

Glu Asn Trp Thr Pro Pro Lys Ala Glu Met Asp Asp Leu Lys Leu Tyr 565 570 575

Leu Glu Thr His Leu Ser Phe Lys Leu Asn Val Asp Ser His Cys Ala 580 585 590

Leu Lys Glu Ala Val Glu Glu Glu Gly His Gin Leu Leu Glu Leu He 595 600 605

Ala Ser His Lys Ala Gly Leu Lys Asp Met Leu Arg Met He Ala Ser 610 615 620

Gin Trp Lys Glu Leu Gin Arg Gin He Lys Arg Gin His Ser Trp He 625 630 635 640

Leu Arg Ala Leu Asp Thr He Lys Ala Glu He Leu Ala Thr Asp Val 645 650 655

Ser Val Glu Asp Glu Glu Gly Thr Gly Ser Pro Lys Ala Glu Val Gin 660 665 670

Leu Cys Tyr Leu Glu Ala Gin Arg Asp Ala Val Glu Gin Met Ser Leu 675 680 685

Lys Leu Tyr Ser Glu Gin Tyr Thr Ser Ser Ser Lys Arg Lys Glu Glu 690 695 700

Phe Ala Asp Met Ser Lys Val His Ser Val Gly Ser Asn Gly Leu Leu 705 710 715 720

Asp Phe Asp Ser Glu Tyr Gin Glu Leu Trp Asp Trp Leu He Asp Met 725 730 735

Glu Ser Leu Val Met Asp Ser His Asp Leu Met Met Ser Glu Glu Gin 740 745 750

Gin Gin His Leu Tyr Lys Val Arg Ala Thr Leu Pro Ala Phe Thr Leu 755 760 765

Leu Trp Lys He 770

(2) INFORMATION FOR SEQ ID NO: 19:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 3442 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: CGCACAGCAT GCCTGGCTGA GAGCTTGAAA CAGAGTTCTG CAGAAAAACT GTAAAGATCC 60 CGAGACATTT CCCTGACTCT TGAGATACTG ACTGGAAGAT AGACTGTTTT GTTCCACCTG 120 ATTGTATGGG AGAAATTTTT GACCTTAGAA AGTGGAAATG AGGTTGCTAT GGAAACTGGT 180 AATTCTGCTG CCACTCATAA ACTCTTCTGC AGGTGATGGT CTTTTAAGCC GTCCTATTTT 240 TACTCAGGAG CCACATGATG TCATTTTTCC TTTGGATTTA TCAAAATCTG AGGTCATCCT 300 GAATTGTGCT GCTAACGGTT ACCCTTCGCC TCATTATAGG TGGAAGCAAA ATGGCACAGA 360 CATTGATTTT ACTATGAGTT ATCACTACAG GTTGGATGGA GGCAGTCTTG CAATCAATAG 420 CCCCCACACA GATCAAGATA TTGGCATGTA CCAGTGCCTG GCCACCAATC TTCTGGGGAC 480 AATTCTGAGT CGGAAGGCAA AGCTCCAATT TGCATATATT GAAGACTTTG AAACTAAAAC 540 AAGAAGCACA GTATCTGTCC GAGAAGGTCA AGGTGTGGTG CTTCTCTGTG GCCCACCGCC 600 ACATTTTGGA GATTTATCTT ATGCATGGAC CTTCAATGAT AACCCCTTAT ACGTCCAAGA 660 GGACAATAGG CGATTTGTAT CTCAAGAGAC GGGAAACTTG TACATTGCCA AAGTGGAACC 720 ATCAGATGTG GGCAACTACA CTTGCTTTAT AACTAACAAA GAGGCCCAGA GAAGTGTTCA 780 AGGTCCACCC ACTCCATTAG TGCAGCGCAC TGATGGTGTG ATGGGGGAAT ATGAACCAAA 840 GATTGAAGTG CGTTTTCCTG AAACTATACA AGCTGCAAAG GATTCATCTG TAAAACTGGA 900 ATGTTTTGCC CTTGGAAATC CAGTCCCCGA TATTAGTTGG AGAAGGTTGG ACGGGAGCCC 960

GTTGCCAGGG AAAGTCAAGT ACAGCAAATC CCAAGCTATC CTTGAAATCC CGAACTTCCA 1020

ACAAGAAGAT GAAGGCTTTT ATGAGTGCAT TGCAAGCAAC CTTCGAGGAA GAAACCTTGC 1080

AAAGGGTCAA CTCATTTTTT ATGCTCCTCC AGAATGGGAA CAGAAAATCC AAAATACACA 1140

CCTCTCTATC TATGACAACT TGCTCTGGGA ATGTAAAGCT AGTGGAAAGC CAAACCCTTG 1200

GTATACATGG TTAAAAAATG GTGAACGACT CAACCCAGAG GAGAGAATTC AAATAGAAAA 1260

TGGGACACTC ATCATAACGA TGCTGAATGT GTCAGATTCT GGTGTGTACC AATGTGCTGC 1320

AGAAAACAAA TATCAGATAA TTTATGCAAA TGCTGAATTG AGAGTTTTAG CCTCAGCTCC 1380

AGATTTCTCC AAAAGTCCAG TTAAAAAAAA GTCTTTTGTT CAAGTTGGTG GGGATATTGT 1440

TATCGGATGC AAACCAAATG CTTTTCCCAG GGCAGCTATC TCTTGGAAAA GAGGAACGGA 1500

GACCCTTAGA CAAAGCAAAA GAATATTTCT CTTGGAGGAT GGCAGCCTCA AGATATATAA 1560

TATTACCAGG TCAGATGCTG GATCATATAC ATGCATAGCC ACAAATCAGT TTGGCACTGC 1620

AAAGAACACT GGCAGCCTCA TTGTAAAAGA GAGAACTGTC ATTACCGTCC CACCTTCCAA 1680

AATGGATGTT ACAGTTGGCG AGAGTATAGT GCTACCATGC CAGGTGTCCC ATGACCCCTC 1740 CATTGAAGTG GTATTTGTAT GGTTTTTCAA TGGAGATGTC ATAGACTTAA AAAAAGGAGT 1800

GGCTCATTTT GAAAGGATTG GAGGAGAATC TGTTGGGGAT TTGATGATAA GGAATATTCA 1860

GTTACATCAT TCAGGAAAAT ATCTCTGCAC AGTACAAACA ACCCTAGAAA GTTTATCTGC 1920

AGTAGCCGAT ATCATTGTTA GAGGTCCACC AGGTCCTCCT GAGGATGTGC AAGTGGAAGA 1980

CATTTCCAGT ACTACTTCTC AACTAAGTTG GAGAGCAGGC CCAGATAATA ACAGTCCCAT 2040

TCAAATATTT ACTATTCAGA CTCGGACACC ATTTTCTGTG GGTTGGCAGG CTGTTGCTAC 2100

AGTTCCAGAA ATTCTCAATG GTAAGACATA CAATGCAACA GTGGTTGGTT TGAGTCCTTG 2160

GGTGGAATAT GAATTTCGTG TTGTTGCCGG CAACAGCATT GGGATTGGAG AACCAAGTGA 2220

ACCATCAGAA TTGTTAAGAA CTAAAGCATC AGTCCCTGTT GTGGCACCAG TAAACATCCA 2280

TGGAGGTGGA GGAAGTCGGT CTGAACTCGT CATTACGTGG GAGTCAATTC CAGAAGAACT 2340

GCAGAATGGG GAGGGATTTG GATATATCAT CATGTTCCGG CCAGTGGGCT CGACAACCTG 2400

GTCCAAGGAG AAAGTATCAT CTGTGGAATC ATCAAGGTTT GTCTACAGAA ATGAAAGCAT 2460

CATCCCACTG TCTCCCTTTG AAGTCAAAGT GGGTGTGTAT AATAATGAAG GAGAAGGATC 2520

CCTGAGTACT GTGACCATTG TCTACTCTGG GGAAGATGAA CCTCAACTGG CCCCAAGGGG 2580

AACTTCTCTC CAGAGTTTTT CTGCTTCTGA AATGGAGGTT TCATGGAATG CTATTGCCTG 2640

GAATAGAAAC ACTGGAAGAG TGCTGGGCTA TGAGGTCTTA TACTGGACAG ATGACTCCAA 2700

AGAATCCATG ATAGGTAAAA TTAGAGTCAG TGGAAATGTC ACAACCAAAA ACATCACGGG 2760

GCTGAAAGCT AATACCATCT ACTTTGCTTC CGTAAGAGCT TACAACACTG CTGGGACAGG 2820

GCCCTCAAGC CCCCCAGTCA ATGTTACCAC CAAAAAGTCT CCTCCAAGCC AACCACCAGC 2880

AAACATTGCC TGGAAGCTGA CAAACTCTAA ATTATGCTTG AACTGGGAGC ATGTAAAAAC 2940

CATGGAAAAT GAGTCTGAAG TTTTGGGGTA CAAGATTCTG TACCGGCAAA ACAGACAGAG 3000

TAAAACTCAT ATTTTGGAAA CAAACAATAC ATCAGCTGAG CTTCTGGTTC CATTTGAAGA 3060

AGACTACTTA ATTGAAATAA GAACAGTCAG TGATGGTGGA GATGGAAGCA GCAGTGAGGA 3120

AATTAGGATT CCAAAAATGT CAAGTTTGAG TTCCAGAGGA ATTCAATTCT TAGAACCTAG 3180

CACCCATTTT CTTTCCATTG TCATTGTGAT TTTTTCACTG TTTTGCTATT CAGCCACTTA 3240

TYTGATGAAT AAAACCATAA ATCTTTGAGA GTTTTTTGAA AGCAAATCAT TCTGTATATA 3300

TGCTCTCCAG CCTCTGACAC AAGATGCGTT CTTAATACAG ACTTGTTTGC AAAGAAAAAA 3360

AAAAGTATAT TATTAAAATC CTGTAAATAT CTATGG ATA TTAATAAAAC AATTTTAAAC 3420 ACTTAAAAAA AAAAAAAAAA AA 3442

(2) INFORMATION FOR SEQ ID NO: 20:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1036 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE-: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 20:

Met Arg Leu Leu Trp Lys Leu Val He Leu Leu Pro Leu He Asn Ser 1 5 10 15

Ser Ala Gly Asp Gly Leu Leu Ser Arg Pro He Phe Thr Gin Glu Pro 20 25 30

His Asp Val He Phe Pro Leu Asp Leu Ser Lys Ser Glu Val He Leu 35 40 45

Asn Cys Ala Ala Asn Gly Tyr Pro Ser Pro His Tyr Arg Trp Lys Gin 50 55 60

Asn Gly Thr Asp He Asp Phe Thr Met Ser Tyr His Tyr Arg Leu Asp 65 70 75 80

Gly Gly Ser Leu Ala He Asn Ser Pro His Thr Asp Gin Asp He Gly 85 90 95

Met Tyr Gin Cys Leu Ala Thr Asn Leu Leu Gly Thr He Leu Ser Arg 100 105 110

Lys Ala Lys Leu Gin Phe Ala Tyr He Glu Asp Phe Glu Thr Lys Thr 115 120 125

Arg Ser Thr Val Ser Val Arg Glu Gly Gin Gly Val Val Leu Leu Cys 130 135 140

Gly Pro Pro Pro His Phe Gly Asp Leu Ser Tyr Ala Trp Thr Phe Asn 145 150 155 160

Asp Asn Pro Leu Tyr Val Gin Glu Asp Asn Arg Arg Phe Val Ser Gin 165 170 175

Glu Thr Gly Asn Leu Tyr He Ala Lys Val Glu Pro Ser Asp Val Gly 180 185 190

Asn Tyr Thr Cys Phe He Thr Asn Lys Glu Ala Gin Arg Ser Val Gin 195 200 205 Gly Pro Pro Thr Pro Leu Val Gin Arg Thr Asp Gly Val Met Gly Glu 210 215 220

Tyr Glu Pro Lys He Glu Val Arg Phe Pro Glu Thr He Gin Ala Ala 225 230 235 240

Lys Asp Ser Ser Val Lys Leu Glu Cys Phe Ala Leu Gly Asn Pro Val 245 250 255

Pro Asp He Ser Trp Arg Arg Leu Asp Gly Ser Pro Leu Pro Gly Lys 260 265 270

Val Lys Tyr Ser Lys Ser Gin Ala He Leu Glu He Pro Asn Phe Gin 275 280 285

Gin Glu Asp Glu Gly Phe Tyr Glu Cys He Ala Ser Asn Leu Arg Gly 290 295 300

Arg Asn Leu Ala Lys Gly Gin Leu He Phe Tyr Ala Pro Pro Glu Trp 305 310 315 320

Glu Gin Lys He Gin Asn Thr His Leu Ser He Tyr Asp Asn Leu Leu 325 330 335

Trp Glu Cys Lys Ala Ser Gly Lys Pro Asn Pro Trp Tyr Thr Trp Leu 340 345 350

Lys Asn Gly Glu Arg Leu Asn Pro Glu Glu Arg He Gin He Glu Asn 355 360 365

Gly Thr Leu He He Thr Met Leu Asn Val Ser Asp Ser Gly Val Tyr 370 375 380

Gin Cys Ala Ala Glu Asn Lys Tyr Gin He He Tyr Ala Asn Ala Glu 385 390 395 400

Leu Arg Val Leu Ala Ser Ala Pro Asp Phe Ser Lys Ser Pro Val Lys 405 410 415

Lys Lys Ser Phe Val Gin Val Gly Gly Asp He Val He Gly Cys Lys 420 425 430

Pro Asn Ala Phe Pro Arg Ala Ala He Ser Trp Lys Arg Gly Thr Glu 435 440 445

Thr Leu Arg Gin Ser Lys Arg He Phe Leu Leu Glu Asp Gly Ser Leu 450 455 460

Lys He Tyr Asn He Thr Arg Ser Asp Ala Gly Ser Tyr Thr Cys He 465 470 475 480

Ala Thr Asn Gin Phe Gly Thr Ala Lys Asn Thr Gly Ser Leu He Val 485 490 495

Lys Glu Arg Thr Val He Thr Val Pro Pro Ser Lys Met Asp Val Thr 500 505 510

Val Gly Glu Ser He Val Leu Pro Cys Gin Val Ser His Asp Pro Ser 515 520 525

He Glu Val Val Phe Val Trp Phe Phe Asn Gly Asp Val He Asp Leu 530 535 540

Lys Lys Gly Val Ala His Phe Glu Arg He Gly Gly Glu Ser Val Gly 545 550 555 560

Asp Leu Met He Arg Asn He Gin Leu His His Ser Gly Lys Tyr Leu 565 570 575

Cys Thr Val Gin Thr Thr Leu Glu Ser Leu Ser Ala Val Ala Asp He 580 585 590

He Val Arg Gly Pro Pro Gly Pro Pro Glu Asp Val Gin Val Glu Asp 595 600 605

He Ser Ser Thr Thr Ser Gin Leu Ser Trp Arg Ala Gly Pro Asp Asn 610 615 620

Asn Ser Pro He Gin He Phe Thr He Gin Thr Arg Thr Pro Phe Ser 625 630 635 640

Val Gly Trp Gin Ala Val Ala Thr Val Pro Glu He Leu Asn Gly Lys 645 650 655

Thr Tyr Asn Ala Thr Val Val Gly Leu Ser Pro Trp Val Glu Tyr Glu 660 665 670

Phe Arg Val Val Ala Gly Asn Ser He Gly He Gly Glu Pro Ser Glu 675 680 685

Pro Ser Glu Leu Leu Arg Thr Lys Ala Ser Val Pro Val Val Ala Pro 690 695 700

Val Asn He His Gly Gly Gly Gly Ser Arg Ser Glu Leu Val He Thr 705 710 715 720

Trp Glu Ser He Pro Glu Glu Leu Gin Asn Gly Glu Gly Phe Gly Tyr 725 730 735

He He Met Phe Arg Pro Val Gly Ser Thr Thr Trp Ser Lys Glu Lys 740 745 750

Val Ser Ser Val Glu Ser Ser Arg Phe Val Tyr Arg Asn Glu Ser He 755 760 765

He Pro Leu Ser Pro Phe Glu Val Lys Val Gly Val Tyr Asn Asn Glu 770 775 780

Gly Glu Gly Ser Leu Ser Thr Val Thr He Val Tyr Ser Gly Glu Asp 785 790 795 800 Glu Pro Gin Leu Ala Pro Arg Gly Thr Ser Leu Gin Ser Phe Ser Ala 805 810 815

Ser Glu Met Glu Val Ser Trp Asn Ala He Ala Trp Asn Arg Asn Thr 820 825 830

Gly Arg Val Leu Gly Tyr Glu Val Leu Tyr Trp Thr Asp Asp Ser Lys 835 840 845

Glu Ser Met He Gly Lys He Arg Val Ser Gly Asn Val Thr Thr Lys 850 855 860

Asn He Thr Gly Leu Lys Ala Asn Thr He Tyr Phe Ala Ser Val Arg 865 870 875 880

Ala Tyr Asn Thr Ala Gly Thr Gly Pro Ser Ser Pro Pro Val Asn Val 885 890 895

Thr Thr Lys Lys Ser Pro Pro Ser Gin Pro Pro Ala Asn He Ala Trp 900 905 910

Lys Leu Thr Asn Ser Lys Leu Cys Leu Asn Trp Glu His Val Lys Thr 915 920 925

Met Glu Asn Glu Ser Glu Val Leu Gly Tyr Lys He Leu Tyr Arg Gin 930 935 940

Asn Arg Gin Ser Lys Thr His He Leu Glu Thr Asn Asn Thr Ser Ala 945 950 955 960

Glu Leu Leu Val Pro Phe Glu Glu Asp Tyr Leu He Glu He Arg Thr 965 970 975

Val Ser Asp Gly Gly Asp Gly Ser Ser Ser Glu Glu He Arg He Pro 980 985 990

Lys Met Ser Ser Leu Ser Ser Arg Gly He Gin Phe Leu Glu Pro Ser 995 1000 1005

Thr His Phe Leu Ser He Val He Val He Phe Ser Leu Phe Cys Tyr 1010 1015 1020

Ser Ala Thr Tyr Leu Met Asn Lys Thr He Asn Leu 1025 1030 1035

(2) INFORMATION FOR SEQ ID NO: 21:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 21: ANTGACTTCTG ATAGGCAACA GTGCCATG 29

(2) INFORMATION FOR SEQ ID NO: 22:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:22: GNGGACTGATG CTTATAAAAT GACATAGC 29

(2) INFORMATION FOR SEQ ID NO: 23:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: CNCACTGCCTG TGAGCACGAT CAAGCCTT 29

(2) INFORMATION FOR SEQ ID NO: 24:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 24: TNGCGCATCTC TTCCTGTTGC TGCTGGTA 29

(2) INFORMATION FOR SEQ ID NO: 25:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 25: ANGCAGCGCTG GAATTTCCTT CATGAGTG 29

(2) INFORMATION FOR SEQ ID NO: 26:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 26: CNTAGTTTCTT GTTCAAGATA ACTGTGGG 29

(2) INFORMATION FOR SEQ ID NO: 27:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 27: GNAGTATCTGT TCCTATTACC ATCTTCCA 29

(2) INFORMATION FOR SEQ ID NO: 28:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 28: TNCTTGGTCAC CTCCAGATGA GTAGCCTG 29

(2) INFORMATION FOR SEQ ID NO: 29:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:29: GNATTGTCTTC CTCACATCCA CCTTTGTC 29

(2) INFORMATION FOR SEQ ID NO: 30:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 30: ANCAGTCTATC TTCCAGTCAG TATCTCAA 29

Claims

What is claimed is:

1. A composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:l;

(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:2;

(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:2 having biological activity;

2. A composition of claim 1 wherein said polynucleotide is operably linked to at least one expression control sequence.

3. A host cell transformed with a composition of claim 2.

4. The host cell of claim 3, wherein said cell is a mammalian cell.

5. A process for producing a protein encoded by a composition of claim 2, which process comprises:

(a) growing a culture of the host cell of claim 3 in a suitable culture medium; and

(b) purifying said protein from the culture.

6. A protein produced according to the process of claim 5.

7. The protein of claim 6 comprising a mature protein.

8. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:2;

(b) the amino acid sequence of SEQ ID NO:2 from amino acid 13 to amino acid 164;

(c) fragments of the amino acid sequence of SEQ ID NO:2; and

(d) the amino acid sequence encoded by the cDNA insert of clone BH272_3 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins.

9. The composition of claim 8, wherein said protein comprises the amino acid sequence of SEQ ID NO:2.

10. The composition of claim 8, wherein said protein comprises the amino acid sequence of SEQ ID NO:2 from amino acid 13 to amino acid 164.

11. The composition of claim 8, further comprising a pharmaceutically acceptable carrier.

12. A method for preventing, treating or ameliorating a medical condition which comprises administering to a mammalian subject a therapeutically effective amount of a composition of claim 11.

13. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:l.

14. A composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:5;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:5 from nucleotide 100 to nucleotide 675;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:5 from nucleotide 208 to nucleotide 489;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CH27_1 deposited under accession number ATCC 98289;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CH27_1 deposited under accession number ATCC 98289;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CH27_1 deposited under accession number ATCC 98289;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CH27_1 deposited under accession number ATCC 98289;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:6;

(j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

15. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(c) fragments of the amino acid sequence of SEQ ID NO:6; and

(d) the amino acid sequence encoded by the cDNA insert of clone CH27_1 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins.

16. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:5.

17. A composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:7;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CI542_2 deposited under accession number ATCC 98289;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CI542_2 deposited under accession number ATCC 98289;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:8;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:8 having biological activity;

(j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above; (k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ; and

18. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:8;

(b) the amino acid sequence of SEQ ID NO:8 from amino acid 170 to amino acid 258;

(c) fragments of the amino acid sequence of SEQ ID NO:8; and

(d) the amino acid sequence encoded by the cDNA insert of clone CI542_2 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins.

19. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:7.

20. A composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:9;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:9 from nucleotide 112 to nucleotide 1212;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:9 from nucleotide 166 to nucleotide 1212;

(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CN483_2 deposited under accession number ATCC 98289; (g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CN483_2 deposited under accession number ATCC 98289;

(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:10 having biological activity;

(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;

(1) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and

(m) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(j).

21. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO: 10;

(b) fragments of the amino acid sequence of SEQ ID NO: 10; and

(c) the amino acid sequence encoded by the cDNA insert of clone CN483_2 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins.

22. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:9.

23. A composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:ll;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 11 from nucleotide 1 to nucleotide 367; (d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone C01224_2 deposited under accession number ATCC 98289;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone C01224_2 deposited under accession number ATCC 98289;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone C01224_2 deposited under accession number ATCC 98289;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone C01224_2 deposited under accession number ATCC 98289;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:12;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 12 having biological activity;

(j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

24. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO: 12;

(b) the amino acid sequence of SEQ ID NO: 12 from amino acid 1 to amino acid 52;

(c) fragments of the amino acid sequence of SEQ ID NO:12; and

(d) the amino acid sequence encoded by the cDNA insert of clone C01224_2 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins.

25. An isolated gene corresponding to the cDNA sequence of SEQ ID NO: 11.

26. A composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:13;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:13 from nucleotide 828 to nucleotide 1111;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 14 having biological activity;

(j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

27. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:14;

(b) fragments of the amino acid sequence of SEQ ID NO: 14; and (c) the amino acid sequence encoded by the cDNA insert of clone CW768_1 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins.

28. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:13.

29. A composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:15;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:15 from nucleotide 116 to nucleotide 1081;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 15 from nucleotide 509 to nucleotide 860;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CZ268_1 deposited under accession number ATCC 98289;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:16 having biological activity;

(j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

30. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO: 16;

(b) the amino acid sequence of SEQ ID NO:16 from amino acid 153 to amino acid 248;

(c) fragments of the amino acid sequence of SEQ ID NO: 16; and

(d) the amino acid sequence encoded by the cDNA insert of clone CZ268_1 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins.

31. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:15.

32. A composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:17;

(a)-(g) above;

33. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO: 18;

(b) the amino acid sequence of SEQ ID NO: 18 from amino acid 19 to amino acid 125;

(c) fragments of the amino acid sequence of SEQ ID NO:18; and

(d) the amino acid sequence encoded by the cDNA insert of clone DH1308_1 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins.

34. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:17.

35. A composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:19;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:19 from nucleotide 158 to nucleotide 3268;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:19 from nucleotide 215 to nucleotide 3268;

(d) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:19 from nucleotide 55 to nucleotide 379;

(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone DL185_1 deposited under accession number ATCC 98289; (g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone DL185_1 deposited under accession number ATCC 98289;

36. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:20;

(b) the amino acid sequence of SEQ ID NO:20 from amino acid 1 to amino acid 74;

(c) fragments of the amino acid sequence of SEQ ID NO:20; and

(d) the amino acid sequence encoded by the cDNA insert of clone DL185_1 deposited under accession number ATCC 98289; the protein being substantially free from other mammalian proteins.

37. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:19.