WO1993023536A1

WO1993023536A1 - Sub-units of nmda receptors, process for producing the same and their use

Info

Publication number: WO1993023536A1
Application number: PCT/EP1993/001169
Authority: WO
Inventors: Alfred Bach; Peter H. Seeburg; Hannah Monyer; Anne Herb
Original assignee: Basf Aktiengesellschaft
Priority date: 1992-05-16
Filing date: 1993-05-12
Publication date: 1993-11-25
Also published as: MX9302841A; DE4216321A1

Abstract

New sub-units of NMDA receptors and the DNA sequences that code for them are disclosed, as well as a process for producing DNA sequences and receptors and a process for identifying functional ligands for said receptor.

Description

Subunits of NDA receptors, processes for their preparation and their use

description

The invention relates to 3 new subunits of NMDA receptors, their use and methods for finding functional ligands for these receptors.

Glutamate is the main excitatory neurotransmitter in the central nervous system. At least five different receptor subtype families are involved in glutamatergic neurotransmission: NMDA, Kainat, AMPA / Quisqualat, AP4 and the metabotropic glutamate receptor (TIPS ü, 1990, 126 - 32, Pharmacological Reviews, 40, No 2,1989,143 - 210). Glutamate receptors play an important role in the Ca ^ homeostasis of neuronal cells.

Glutamate is involved in a variety of physiological and pathophysiological processes. In the case of pathophysiological situations, epilepsy, schizophrenia, neurodegenerative diseases and ischemic states of different origins are to be mentioned in particular. In the physiological processes, the cognitive functions such as B. Learn to think. As a result, receptors for glutamate are interesting targets for pharmaceuticals for the treatment of the abovementioned. pathological conditions.

Some primary units of AMPA and kainate receptors, one subunit of an NMDA receptor (NR1) and some metabotropic receptors (Nature 112, 643, 1989, Science 231, 556, 1990, Neuron £, 169, 1992) have so far been elucidated in their primary structure ).

New subunits of rat NMDA receptors have now been found, as well as DNA sequences which code for such subunits.

SEQ ID NO 1 shows the cDNA sequence of NR2A and the polypeptide sequence derived therefrom (SEQ ID NO 2); in SEQ ID NO 3 the cDNA sequence of NR2B, in SEQ ID NO 4 the derived polypeptide sequence, in SEQ ID NO 5 the cDNA sequence of NR2C, in SEQ ID NO 6 the derived polypeptide sequence.

Other suitable DNA sequences are those which have a different nucleotide sequence than that listed in ^' SEQ ID NO 1, 3 or 5, but which, owing to the degeneracy of the genetic code for those listed in SEQ ID NO 1, 3 or 5 Encode polypeptide chain or parts thereof. Such DNA sequences are also suitable gnet, which code for NMDA receptor subunits and which hybridi under standard conditions with the nucleotide sequence shown in SEQ ID NO 1, 3 or 5 or with a nucleotide sequence which codes for the protein shown in SEQ ID NO 2, 4 or 6 - sieren. Standard conditions are understood to mean, for example, temperatures between 42 and 58 ° C. in an aqueous buffer solution with a concentration between 0.1 and 1 x SSC (1 x SSC: 0.15M NaCl, 15mM sodium nitrate pH 7.2). The experimental conditions for DNA hybridization are described in textbooks of genetic engineering, for example in Sambrook et al., "Molecular Cloning", Cold Spring Harbor Laboratory, 1989.

Furthermore, genetic engineering manufacturing processes for these subunits have been found. It was also found that the DNA sequences coding for these receptor subunits can be used to find functional ligands for these receptors. The invention furthermore relates to methods for identifying functional ligands for NMDA receptors, which are characterized in that cells with sequences which code for NMDA receptor subunits are transfected, the membranes of these cells are isolated and with these membranes customary receptor binding experiments carries out. Another method according to the invention for identifying functional ligands for NMDA receptors is characterized in that changes in the intracellular Ca ⁺⁺ concentration are made in cells which have been transfected with DNA sequences which code for NMDA receptor subunits detected by fluorimetric methods after ligand binding.

The new polypeptides and DNA sequences can be genetically engineered using known methods. In this way, one can isolate mRNA from brain tissue and translate it into double-stranded cDNA. This cDNA can be used as a template for the polymerase chain reaction. By using specific primers, the corresponding cDNA a can be plotted under suitable reaction conditions. By using suitable primers, the amplified cDNA can be sequenced without prior cloning. The double-stranded cDNA can also be used in λ vectors e.g. B. λ gt 10 or λ ZAP can be integrated to generate a brain-specific cDNA bank. Such a cDNA bank can be searched with radiolabelled DNA or RNA probes in order to identify clones which have homology with the hybridization sample. The methods used are described, for example, in "Current Protocols in Molecular Biology" (ed. FM Ausubel et al.) 1989, ISBN 0-471 50338-x (Vol. 1 and 2), for the polymerase chain reactions. tion in Saiki et al. (1985) Science, 230, 1350-54 and Mullis and Faloona (1987) Meth. Enzy ol., 155, 335-350.

The cDNA characterized in this way is easily accessible with the aid of restriction enzymes. The resulting fragments, possibly in conjunction with chemically synthesized oligonucleotides, adapters or gene fragments, can be used to clone the sequences coding for the protein. The incorporation of the gene fragments or synthetic DNA sequences into cloning vectors, eg. B. the commercially available plasmids Ml3mpl8 or Bluescript, is carried out in a known manner. The genes or gene fragments can also be provided with suitable chemically synthesized control regions or control regions isolated from bacteria, phages, eukaryotic cells or their viruses, which enable expression of the proteins in different host systems.

The transformation or transfection of suitable host organisms with hybrid plasmids is also known and described in detail (M. Wigler et al., Cell, 16 (1979), 777-785; FL Graham and AJ van der Eb, Virology, 52 (1973) , 456-467).

For expression in mammalian cells it is possible to use vectors which control the mouse metallothionein, the viral SV40, the gene to be expressed, in this case the cDNA sequences coding for the subunits of NMDA receptors described here. or the cytomegalovirus promoter (J. Page Martin, Gene, 37 (1985), 139-144). Expression is dependent on the presence of the methionine start codon of the gene which codes for these subunits of NMDA receptors. Clones are then isolated which have copies of these vectors as episomes or integrated into the genome. The integration of the foreign gene into a vector which contains the promoter of the cytomegalovirus is particularly advantageous.

Alternatively, cells can be transfected with a suitable vector such that the transient expression of the DNA thus introduced is sufficient for pharmacological characterization of the expressed heterologous polypeptides. Here, too, control of expression by the cytomegalovirus promoter is particularly advantageous.

Furthermore, functional NMDA receptors can be produced by transfecting one or more DNA sequences from the group NR2A, NR2B and NR2C together with the DNA sequence for NR1 (Nature, 354, 31 (1991)) into cells. In this way, NMDA receptors with different subunits can be obtained. In connection with prokaryotic sequences which code for replication in bacterial cells and antibiotic resistance, the use of "shuttle" vectors is well suited. The plasmid is first constructed and propagated in bacterial cells; then the conversion into the eukaryotic cells takes place, eg. B. in the embryonic human kidney cell line HEK 293.

Other cell systems, e.g. B. yeast and other fungi, insect cells and animal and human cells such. B. CHO, COS and L cells can be used in conjunction with suitable expression vectors to express the cloned cDNA.

The eukaryotic expression systems have the advantage that they are able to express their products effectively and mostly in their native form. They also have the ability to modify their products post-translationally.

The expressed receptor proteins can be solubilized by detergents and purified by affinity chromatography using known methods. After crystallization and X-ray structure analysis or other physical methods such as NMR or scanning tunneling microscopy, the pure polypeptide can be used to clarify the spatial structure of the ligand binding site.

After appropriate purification, the expressed receptor proteins can also serve as antigens for the generation of polyclonal or monoclonal antibodies. These antibodies in turn can be used for diagnostic purposes if necessary. Another application for such antibodies is to use them as an aid to rational drug design. For example, the receptor-specific antibodies can be used as antigens for the generation of anti-idiotypic antibodies. Such antibodies can represent an image of the receptor for defined areas and can be used for screening for specific receptor ligands or for rational drug design.

Receptor-expressing cell lines represent an important tool in the screening for specific receptor ligands. For this purpose, the membranes of these cells can be used for the receptor binding test. Information about the mode of action (agonism / antagonism) of a receptor ligand can be obtained by providing cells which have been transfected with a DNA sequence according to the invention with a suitable reporter system. Suitable reporter systems are those in which a promoter which is linked by the signal transduction pathway (second measured senger) is functionally linked to a gene for an easily detectable product such as luciferase. Such reporter systems are for example from Science 252, 1424 (1991) or Proc. Natl. Acad. Be. USA 88, 5061 (1991). A suitable promoter, which is regulated, for example, by the intracellular Ca ⁺⁺ concentration, is that of the fos gene. It is also possible to change the intracellular Ca ⁺⁺ concentration with fluorescent dyes such. B. FURA 2AM) directly.

Furthermore, the current flow through the cell membrane can be measured as a function of the ligand binding.

Due to the degeneration of the genetic code, it is possible to use different DNA sequences than described here, e.g. B. use chemically synthesized genes with different DNA sequences for the expression of the described subunits of NMDA receptors of the rat.

With the aid of the invention it becomes possible to identify and characterize substances which bind to the receptor described here and act there agonistically or antagonistically.

Further refinements of the invention are described in more detail in the examples.

For genetic engineering methods such. B. on the manual of Sambrook et al. 'Molecular Cloning ", Cold Spring Harbor Laboratory, 1989, or" DNA cloning ", Vol. I to III, IRI Press 1985 to 1987, publisher D.M. Glover.

example 1

Isolation of cDNA molecules which code for NMDA receptor subunits of the rat

0.5 g of a rat cerebrum was digested in 6 M guanidinium thiocyanate, 5 mM sodium citrate (pH 7.0), 0.1 M 2-mercaptoethanol, 0.5% sarcosyl in the ULTRA-TURRAX. Coarse cell debris was centrifuged off at 3,000 rpm in the Sorvall SS34 rotor. The RNA was separated by centrifugation through a 5.7 M CsCl cushion at 45,000 rpm for 12 hours. The polyA ⁺ -containing RNA fraction was then separated by affinity chromatography on oligo (dT) cellulose.

Using the enzyme AMV reverse transcriptase and oligo (dT) ι _i ₈ as starters, the polyA ⁺ RNA was rewritten into single-stranded cDNA. The second strand was synthesized using E. coli DNA polymerase I. An EcoRI adapter with the following sequence was attached to the double-stranded cDNA with the aid of the enzyme T4-DNA ligase: 5'AATT CCATGGATGCATGC 3 '(SEQ ID NO 7). The commercially available phage vector λ gt 10 was linearized with the restriction enzyme EcoRI. Phage DNA and cDNA were ligated together and packaged with infectious phages using a commercially available packaging extract (Promega). The recombinant phages were plated with E. coli C 600 Hfl on NZYDT plates (GIBCO / BRL) and incubated at 37 ° C. overnight. The cDNA library thus obtained contained 2x10 ⁶ independent clones. After amplification of the cDNA library in accordance with conventional methods, 500,000 phages were plated with C 600 Hfl cells. The phages were transferred to nitrocellulose filters, lysed with 0.5 N NaOH / 1.5 M NaCl and the denatured DNA was firmly bound to the filter by baking at 80 ° C. for 2 hours. The filters were in

6 x SET buffer (1 x SET = 0.15 M NaCl, 15 mM Tris / HCl, pH 7.4, 1 mM EDTA), 0.1% SDS and 5 x Denhardt's solution (100 x Denhardt = 1 g Ficoll , 1 g polyvinylpyrrolidone, 1 g BSA per 50 ml) prehybridized at 68 ° C for 4 h.

Hybridization was carried out with a nick-translated cDNA sample, which was prepared as follows:

Two degenerate oligonucleotides were derived from peptide regions that are highly conserved within known glutantate receptor subunits.

These oligonucleotides had the following sequence:

CA) 5'-GCGAATTCTGGAA (C, T) GG (C, A) TGATGGG (G, A, T, C) GA-3 '(SEQ ID NO 8)

(B) 5'-GCGGTACCAA (A, G) GC (A, T, G) CCA (A, G) (A, G) TT (A, T, G) GC (A, T). GT (A, G) T-3 '(SEQ ID NO 9)

They correspond to the peptide regions WNGHHGEI (SEQ ID NO 10) (approx. 60 amino acid residues from the N-terminal end of the 1st transmembrane region) and YTANLAAF (SEQ ID NO 11) (at the C-terminal end of the 3rd transmembrane domain). These oligonucleotides were used as primers in the polymerase chain reaction (PCR). 20 ng double-stranded rat brain cDNA (see above) was used as template. The primer concentration was 1 mM, the reaction volume 50 mM. The reaction conditions were: 10 mM Tris / HCl, pH 8.7, 50 mM KC1, 2.5 MgCl ₂ , each 0.2 mM dATP, dCTP, dGTP and dTTP. 1 unit of Thermus aquaticus DNA polymerase (Perkin-Elmer / Cetus) was added. 35 cycles were carried out with the following temperature profile: 94 ° C. for 30 seconds, 55 ° C. for 40 seconds, 72 ° C. for 1 minute. The thermal cycler 9600 from Perkin-Elmer. The amplified DNA (approx. 500 base pairs) was cloned into the vector pCR 1000 and multiplied in accordance with the manufacturer's instructions using the TA Cloning Kit (Invitrogen).

For radioactive labeling, the cloned-in application product was released again with the aid of the restriction enzymes Eco Rl and Hind III by electrophoresis in an agarose gel and separated from the vector using the Geneclean II kit (from the gel). Niek translation was carried out using the Boehringer Mannheim nick translation kit in accordance with the manufacturer's instructions. Α [ ³ P] -dCTP from Amersham Buchler with a specific activity of> 3000 Ci / mmol was used. The nick-translated DNA had a specific activity of 5 x 10 ⁸ dpm / μg DNA.

The nitrocellulose filters to which the phage DNA adhered (see above) were in a solution containing 5 x SET, 0.1% SDS, 30% formamide, 5 x Denhardt's, 10% dextran sulfate and the nick transla - Contained sample with a concentration of 10 ⁶ dpm / ml, incubated for 16 hours at 42 ° C with gentle shaking. They were then washed four times in 2 x SET / 0.1% SDS at 42 ° C. for 20 minutes, dried and exposed to an X-ray film. Clones that gave a radioactive response when "screened" were isolated and grown to obtain the appropriate phage DNA.

Phage DNA was prepared by incubating the purified phages with proteinase K (ad 60 μg / ml) at 55 ° C. for 1 h and then phenol / chloroform extraction. After adding 3 volumes of ethanol (-20 ° C.), the phage DNA precipitated and was transferred into 70% ethanol using a sterile injection needle, washed and briefly sedimented. After briefly air drying, the pellet was suspended in TE buffer.

Example 2

Production of single-stranded DNA coding for subunits of the rat NMDA receptor

The starting point was the phage DNA sequences described in Example 1. They were each individually cut preparatively with the restriction enzyme Eco Rl. The Eco R1 fragments containing the cDNA insertions were eluted electrophoretically from the gel. 30 ng of each of these fragments were ligated at 4 ° C. for 12 h with 100 ng of the Eco RI cut, commercially available cloning vector M13mpl8 or M13mpl9. The volume of the Ligation batch was 10 ul. The ligation was terminated by heating to 80 ° C for 5 minutes.

1/10 volume of each ligation batch was used to transform 100 μl competent JM 101 cells. After completion of the transformation, 60 μl of 0.2 M IPTG solution and 120 μl of XGal (20 mg / ml) were added to the transformation mixture. This An¬ was set plated in top agar on NZYDT NZYDT-agar plates with 200 ul JM101 cells (ODεoo = D. The ^"medium NZYDT is com- mercially available (GIBCO-BRL). Clones cDNA insertions ent held, The sequence analysis of this class showed that 3 new subunits of a receptor were found which have homology with an NMDA receptor subunit of the NMDA receptor.

The clones which coded for the new NMDA receptor subunits were designated NR2A, NR2B and NR2C. Their DNA and the protein sequences derived from them are shown in SEQ ID NO 1 to 6.

SEQ ID NO 1: NR2A cDNA sequence SEQ ID NO 2: NR2A polypeptide sequence SEQ ID NO 3: NR2B cDNA sequence SEQ ID NO 4: NR2B polypeptide sequence SEQ ID NO 5: NR2C cDNA sequence SEQ ID NO 6: NR2C polypeptide sequence

Example 3

Transient expression of the cloned rat receptor genes in HEK 293 cells

Unless otherwise described, the cell culture was carried out according to Lindl and Bauer, "Zeil and tissue culture", Gustav Fischer Verlag.

Double-stranded M 13 mp 18 DNA, which contained NMDA receptor-encoding cDNA from Example 2 as an insert, was cleaved with the enzymes EcoRI and Hindin. The resulting DNA fragment with overhanging ends coding for the receptor was treated using the enzymes T ₄ -DNA polymerase and Klenow fragment of the E. coli DNA polymerase according to standard conditions (Current Protocols in Molecular Biology see above) to generate smooth ends. The commercially available linkers (Invitrogen) with the sequence 5'-CTTAGAGCACA-3 '3'-GAATCTC-5' (SEQ ID NO 12) were then ligated to this fragment. That with these linkers DNA fragment was ligated under standard conditions into the commercially available BstXI cut vectors pCDM8 and RC / CMV (Invitrogen). The resulting recombinant plasmids were propagated in a known manner. 5

HEK 293 cells were cultivated under standard conditions in a 10 cm cell culture dish up to a cell number of 7 to 8 × 10 ⁶ cells. After trypsinization, the cells were diluted 1: 3 in MEM medium (Gibco), which contained 2.2 g / 1 NaHC0, and sown again in 10 10 cm petri dishes. Thereafter, the cells were cultured for 40 to 48 h at 37 ° C ^'.

The DNA to be transfected was prepared as follows: 20 43 g of the DNA solution (1 mg / ml), purified over a CsCl gradient, were mixed with 15 437 μl H 0, then 62.5 μl 2 M CaCl was added and finally 500 µl BBS. Ca ⁺⁺ precipitates formed within 10 min at room temperature.

The solution was placed on a 10 cm cell culture dish with the HEK 293 cells cultured according to the above instructions. After thorough mixing, the cells were cultivated for 15 to 20 h in an incubator at 37 ° C./3% CO ₂ . 5 ml of serum-free medium were then carefully added. After removing all of the medium and repeating the washing process with 5 ml of serum-free medium, 10 ml of complete medium were added to the cells. After 48 h of incubation with 5% CO, the cells could be used for pharmacological and electrophysiological studies.

Alternatively, the DNA was also introduced into the cells 30 in a liposome-mediated manner. Lipofectin from GIBCO-BRL was used in accordance with the manufacturer's instructions.

Example 4

35 Expression of NMDA receptor subunits in oocytes

To produce cRNA, the corresponding cDNA, which codes for the NMDA receptor subunits, was cloned into the plasmid Bluescript (Stratagene), which had been cleaved with EcoRI, according to standard protocols. The cDNAs were released from the λ phage DNA described above by cleavage with the restriction enzyme EcoRI and purified electrophoretically.

After the Bluescript clones, which code for subunits 45 of the NMDA receptor, had been multiplied, plasmid DNA was obtained by standard methods. This plamid DNA was cleaved with the restriction enzyme Not I and used for in vitro transcription. The transcription was started by the T3 or T7 promoter and carried out under standard conditions in accordance with the Stratagene in vitro transcription kit.

To express the receptor subunits, 10 ng cRNA of (NR2A, NR2B or NR2C) were injected either individually or in combination with NRl [Nature, 354, 31 (1991)] into oocytes that had been explanted from the clawed frog Xenopus laevis [C. Meth shackle et al .. Pflügers Arch. 407, 577, ^"(1986)]. The oocytes wur- in the OR-2 (92.5 mM NaCl, 2.5 mM KCl, Na ₂ HP0 ₄ imm, 5 mM HEPES, 1 mM MgCl ₂ , 1 mM CaCl, 0.5 g / 1 polyvinylpyrolidone, pH 7.2 with the addition of 4 μg / ml Zinacef and 100 U / ml Penstrep) at 19 ° C. after 24 hours After the injection, the oocytes were treated with collagenase (type II Sigma) (1 mg / ml in OR-2 for 1 hour.) Electrophysiological recordings were made 2-6 days after the cRNA was injected. "Voltage clamp" configuration used with a TEC 01C A plifier (NPI Electronic, Tarnm, Germany) .. During the electrophysiological measurements, the oocytes were treated with normal Frog ring solution NFR (115 mM NaCl, 2.5 mM KCl, 1 , 8 mM CaCl, 10 mM HEPES, pH 7.2) perfused.

SEQUENCE LOG

(1) GENERAL INFORMATION:

(i) APPLICANT:

(A) NAME: BASF Aktiengesellschaft

(B) STREET: Carl-Bosch-Strasse 38

(C) LOCATION: Ludwigshafen

(E) COUNTRY: Federal Republic of Germany

(F) POSTAL NUMBER: D-6700

(G) TELEPHON: 0621/6048526 (H) TELEFAX: 0621/6043123 (I) TELEX: 1762175170

(ii) APPLICATION TITLE: Subunits of NMDA receptors, processes for their preparation and their use (iii) NUMBER OF SEQUENCES: 12

(iv) COMPUTER READABLE FORM:

(A) DISK: Floppy disk

(B) COMPUTER: IBM PC compatible

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

(D) SOFTWARE: Patentin Release # 1.0, Version # 1.25 (EPA)

(2) INFORMATION ABOUT SEQ ID NO: 1:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 5146 base pairs

(B) ART: nucleic acid

(C) STRAND FORM: Single

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

(iii) HYPOTHETICAL: NO (iii) ANTISENSE: NO (vi) ORIGINAL ORIGIN:

(A) ORGANISM: Rattus norvegicus (F) FABRIC TYPE: Brain (ix) CHARACTERISTICS:

(A) NAME / KEY: CDS

(B) LOCATION: 432..4826

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

GCAGGAGCAG GACTGCGAGA GGGTGTCCAG GAGCGCTCAG AGGACCGGCA GTCGCTGTCC

GGAGTGGAAC AGAAAGCTGA GCCAGGGCTC TAGAAGAGAG GCTCCTGAGG TGCTGTGCCT 1

GAGCATGGGG CTGGATGAGG TCTGAGAGTC GCGGCGGCAG CAATCAGCCC TGGAGATGAC 1

CAGGGGTGGC CACTGCTGAG AACTATGTGG AGAGAGGCTG CGAGCCCTGC TGCAGAGCCT 2

CCGGCTGGGA TAGCCGCCCC CCGTGGGGGT GATGCGGACA GCGCGGGACA GCCAGGGGAG 3

CGCGCGGGGG CCGCAGCATG CGGGAACCCG CTAAACCTGG TGGCTGCTGA GGCGGCCGAG 3

ATGCTCGTGC GCGCAGCACG CCCCATTGCA TCCTCCACCT TCTCCGGCTA CAGGGACCCT 4

AAGTGGCGAC C ATG GGC AGA TTG GGC TAC TGG ACC TTG CTG GTA TTG CCG 4 Met Gly Arg Leu Gly Tyr Trp Thr Leu Leu Val Leu Pro 1 5 10 GCC CTT CTG GTC TGG CGC GAT CCG GCG CAG AAC GCG GCG GCG GAG AAG 51 Ala Leu Leu Val Trp Arg Asp Pro Ala Gin Asn Ala Ala Ala Glu Lys

15 20 25

GGT CCT CCA GCG CTG AAC ATT GCG GTG CTG CTG GGT CAC AGC CAC GAC 56 Gly Pro Pro Ala Leu Asn Ile Ala Val Leu Leu Gly His Ser His Asp 30 35 40 45

GTG ACA GAA CGC GAA CTT CGA AAT CTG TGG GGC CCA GAG CAG GCA ACC 61 Val Thr Glu Arg Glu Leu Arg Asn Leu Trp Gly Pro Glu Gin Ala Thr

50 55 60

GGC TTG CCC CTG GAT GTG AAC GTG GTG GCG TTA TTG ATG AAC CGC ACT 66 Gly Leu Pro Leu Asp Val Asn Val Val Ala Leu Leu Met Asn Arg Thr

65 70 75

GAC CCT AAG AGC CTC ATC ACG CAT GTG TGC GAC CTC ATG TCC GGG GCG 71 Asp Pro Lys Ser Leu Ile Thr His Val Cys Asp Leu Met Ser Gly Ala

80 85 90

CGC ATC CAC GGC TTG GTG TTT GGA GAT GAC ACG GAC CAG GAG GCT GTG 75 Arg Ile His Gly Leu Val Phe Gly Asp Asp Thr Asp Gin Glu Ala Val

95 100 105

GCC CAG ATG CTG GAT TTT ATC TCC TCA CAG ACT TTT ATC CCC ATC TTG 80 Ala Gin Met Leu Asp Phe Ile Ser Ser Gin Thr Phe Ile Pro Ile Leu 110 115 120 125

GGC ATT CAT GGG GGT GCA TCT ATG ATC ATG GCT GAC AAG GAT CCG ACA 85 Gly Ile His Gly Gly Ala Ser Met Ile Met Ala Asp Lys Asp Pro Thr

130 135 140

TCC ACG TTC TTC CAG TTT GGA GCC TCC ATC CAG CAG CAA GCC ACA GTT 90 Ser Thr Phe Phe Gin Phe Gly Ala Ser Ile Gin Gin Gin Ala Thr Val

145 150 155

ATG CTG AAG ATC ATG CAG GAC TAC GAC TGG CAC GTC TTC TCC CTG GTC 95 Met Leu Lys Ile Met Gin Asp Tyr Asp Trp His Val Phe Ser Leu Val

160 165 170

ACC ACC ATC TTC CCT GGC TAC CGA GAC TTC ATC AGC TTT ATC AAG ACA 99 Thr Thr Ile Phe Pro Gly Tyr Arg Asp Phe Ile Ser Phe Ile Lys Thr

175 180 185

ACA GTG GAC AAC AGC TTT GTG GGC TGG GAT ÄTG CAG AAC GTG ATC ACA 104 Thr Val Asp Asn Ser Phe Val Gly Trp Asp Met Gin Asn Val Ile Thr 190 195 200 205

CTG GAC ACC TCC TTC GAG GAC GCC AAG ACG CAG GTC CAG CTG AAG AAG 109 Leu Asp Thr Ser Phe Glu Asp Ala Lys Thr Gin Val Gin Leu Lys Lys

210 215 220

ATC CAT TCT TCT GTC ATC CTG CTC TAC TGC TCC AAA GAC GAG GCT GTC 114 Ile His Ser Ser Val Ile Leu Leu Tyr Cys Ser Lys Asp Glu Ala Val

225 230 235

CTC ATC CTG AGC GAG GCT CGC TCC CTC GGC CTC ACT GGC TAT GAT TTC 119 Leu Ile Leu Ser Glu Ala Arg Ser Leu Gly Leu Thr Gly Tyr Asp Phe 240 245 250 TTC TGG ATT GTC CCC AGT TTG GTG TCT GGG AAC ACA GAG CTC ATC CCC 123 Phe Trp Ile Val Pro Ser Leu Val Ser Gly Asn Thr Glu Leu Ile Pro

255 260 265

AAA GAG TTT CCA TCA GGT CTC ATT TCA GTC TCT TAT GAC GAC TGG GAC 128 Lys Glu Phe Pro Ser Gly Leu Ile Ser Val Ser Tyr Asp Asp Trp Asp 270 275 280 285

TAC AGC CTG GAG GCA AGA GTG AGA GAC GGT CTT GGG ATC TTA ACC ACT 133 Tyr Ser Leu Glu Ala Arg Val Arg Asp Gly Leu Gly Ile Leu Thr Thr

290 295 300

GCC GCA TCC TCC ATG TTG GAG AAG TTC TCC TAC ATT CCT GAG GCC AAG 138 Ala Ala Ser Ser Met Leu Glu Lys Phe Ser Tyr Ile Pro Glu Ala Lys

305 310 315

GCC AGC TGC TAT GGG CAG GCA GAG AAG CCA GAG ACC CCG CTA CAC ACC 143 Ala Ser Cys Tyr Gly Gin Ala Glu Lys Pro Glu Thr Pro Leu His Thr

320 325 330

CTG CAC CAA TTC ATG GTC AAT GTG ACT TGG GAT GGC AAG GAC TTG TCC 147 Leu His Gin Phe Met Val Asn Val Thr Trp Asp Gly Lys Asp Leu Ser

335 340 345

TTC ACT GAG GAA GGT TAC CAG GTG CAC CCC AGG CTT GTG GTG ATC GTG 152 Phe Thr Glü Glu Gly Tyr Gin Val His Pro Arg Leu Val Val Ile Val 350 355 360 365

CTG AAC AAG GAC CGG GAG TGG GAA AAG GTG GGC AAG TGG GAG AAC CAG 157 Leu Asn Lys Asp Arg Glu Trp Glu Lys Val Gly Lys Trp Glu Asn Gin

370 375 380

ACG CTG AGC CTG AGG CAC GCT GTG TGG CCA AGG TAC AAG TCC TTT TCT 162 Thr Leu Ser Leu Arg His Ala Val Trp Pro Arg Tyr Lys Ser Phe Ser

385 390 395

GAC TGC GAG CCA GAT GAC AAC CAC CTC AGC ATT GTC ACC TTG GAG GAA 167 Asp Cys Glu Pro Asp Asp Asn His Leu Ser ile Val Thr Leu Glu Glu

400 405 410

GCC CCC TTC GTC ATC GTA GAG GAC ATA GAC CCC CTG ACT GAG ACC TGT 171 Ala Pro Phe Val Ile Val Glu Asp Ile Asp Pro Leu Thr Glu Thr Cys

415 420 425

GTG AGG AAC ACG GTG CCC TGT CGG AAG TTC GTC AAG ATC AAC AAT TCA 176 Val Arg Asn Thr Val Pro Cys Arg Lys Phe Val Lys Ile Asn Asn Ser 430 435 440 445

ACC AAC GAA GGG ATG AAT GTG AAG AAA TGC TGC AAG GGG TTC TGC ATC 18 Thr Asn Glu Gly Met Asn Val Lys Lys Cys Cys Lys Gly Phe Cys Ile

450 455 460

GAC ATC CTC AAG AAG CTG TCC AGA ACT GTG AAG TTC ACC TAT GAC CTC 18 Asp Ile Leu Lys Lys Leu Ser Arg Thr Val Lys Phe Thr Tyr Asp Leu

465 470 475

TAC CTG GTG ACC AAT GGG AAG CAT GGG AAA AAG GTT AAC AAT GTG TGG 19 Tyr Leu Val Thr Asn Gly Lys His Gly Lys Lys Val Asn Asn Val Trp 480 485 490 AAT GGA ATG ATA GGT GAA GTG GTC TAT CAA CGA GCA GTC ATG GCT GTG 1958 Asn Gly Met Ile Gly Glu Val Val Tyr Gin Arg Ala Val Met Ala Val

495 500 505

GGC TCA CTC ACC ATC AAT GAG GAG CGT TCG GAA GTG GTG GAC TTC TCG 2006 Gly Ser Leu Thr Ile Asn Glu Glu Arg Ser Glu Val Val Asp Phe Ser

510 515 520 525

GTG CCC TTC GTG GAG ACA GGA ATC AGC GTC ATG GTC TCC AGG AGT AAT 2054 Val Pro Phe Val Glu Thr Gly Ile Ser Val Met Val Ser Arg Ser Asn

530 535 540

GGC ACT GTC TCC CCT TCT GCT TTC CTC GAA CCC TTC AGT GCC TCC GTC 2102 Gly Thr Val Ser Pro Ser Ala Phe Leu Glu Pro Phe Ser Ala Ser Val

545 550 555

TGG GTG ATG ATG TTT GTG ATG CTG CTC ATC GTC TCA GCC ATT GCT GTC 2150 Trp Val Met Met Phe Val Met Leu Leu Ile Val Ser Ala Ile Ala Val

560 565 570

TTC GTT TTT GAA TAC TTC AGT CCT GTT GGA TAC AAC AGA AAC TTA GCC 2198 Phe Val Phe Glu Tyr Phe Ser Pro Val Gly Tyr Asn Arg Asn Leu Ala

575 580 585

AAA GGG AAA GCT CCC CAC GGG CCT TCT TTT ACT ATT GGA AAA GCT ATA 2246 Lys Gly Lys Ala Pro His Gly Pro Ser Phe Thr Ile Gly Lys Ala Ile 590 595 600 605

TGG CTC CTC TGG GGC CTG GTC TTC AAC AAT TCT GTG CCT GTC CAG AAT 2294 Trp Leu Leu Trp Gly Leu Val Phe Asn Asn Ser Val Pro Val Gin Asn

610 615 620

CCT AAA GGC ACA ACC AGC AAG ATC ATG GTG TCA GTG TGG GCC TTC TTT 2342 Pro Lys Gly Thr Thr Ser Lys Ile Met Val Ser Val Trp Ala Phe Phe

625 630 635

GCT GTC ATC TTC CTG GCC AGT TAC ACA GCC AAC CTG GCT GCC TTC ATG 2390 Ala Val Ile Phe Leu Ala Ser Tyr Thr Ala Asn Leu Ala Ala Phe Met

640 645 650

ATC CAG GAG GAG TTT GTG GAC CAA GTG ACT GGC CTC AGT GAC AAG AAG 2438 Ile Gin Glu Glu Phe Val Asp Gin Val Thr Gly Leu Ser Asp Lys Lys

655 660 665

TTC CAG AGA CCT CAT GAC TAT TCT CCA CCT TTC CGA TTT GGG ACG GTA 2486 Phe Gin Arg Pro His Asp Tyr Ser Pro Pro Phe Arg Phe Gly Thr Val 670 675 680 685

CCC AAT GGA AGT ACA GAG AGG AAT ATT CGT AAC AAC TAC CCC TAT ATG 2534 Pro Asn Gly Ser Thr Glu Arg Asn Ile Arg Asn Asn Tyr Pro Tyr Met

690 695 700

CAC CAG TAC ATG ACC AGA TTC AAC CAG AGG GGA GTG GAG GAT GCC TTG 2582 His Gin Tyr Met Thr Arg Phe Asn Gin Arg Gly Val Glu Asp Ala Leu

705 710 715

GTC AGC TTG AAA ACC GGG AAG TTG GAC GCT TTC ATC TAT GAC GCA GCC 2630 Val Ser Leu Lys Thr Gly Lys Leu Asp Ala Phe Ile Tyr Asp Ala Ala

720 725 730 GTC TTG AAC TAC AAG GCC GGG AGG GAT GAA GGC TGT AAA CTG GTG ACC 267 Val Leu Asn Tyr Lys Ala Gly Arg Asp Glu Gly Cys Lys Leu Val Thr

735 740 745

ATT GGG AGC GGG TAC ATC TTT GCT AGC ACA GGC TAT GGA ATT GCG CTG 272 Ile Gly Ser Gly Tyr Ile Phe Ala Ser Thr Gly Tyr Gly Ile Ala Leu 750 755 760 765

CAG AAG GGC TCA CCC TGG AAG AGG CAG ATT GAC CTC GCT CTG CTC CAG 277 Gin Lys Gly Ser Pro Trp Lys Arg Gin Ile Asp Leu Ala Leu Leu Gin

770 775 780

TTT GTT GGT GAT GGT GAG ATG GAG GAG CTG GAG ACA CTG TGG CTT ACG 282 Phe Val Gly Asp Gly Glu Met Glu Glu Leu Glu Thr Leu Trp Leu Thr

785 790 795

GGC ATC TGC CAC AAC GAG AAG AAT GAG GTG ATG AGT AGC CAG CTG GAC 287 Gly Ile Cys His Asn Glu Lys Asn Glu Val Met Ser Ser Gin Leu Asp

800 805 810

ATC GAT AAC ATG GCG GGC GTG TTC TAC ATG CTG GCT GCA GCC ATG GCC 291 Ile Asp Asn Met Ala Gly Val Phe Tyr Met Leu Ala Ala Ala Meta Ala

815 820 825

CTC AGC CTC ATC ACC TTC ATC TGG GAG CAC CTC TTC TAC TGG AAG CTG 296 Leu Ser Leu Ile Thr Phe Ile Trp Glu His Leu Phe Tyr Trp Lys Leu 830 835 840 845

CGC TTC TGC TTC ACA GGC GTG TGC TCT GAC CGG CCC GGG CTG CTC TTC 301 Arg Phe Cys Phe Thr Gly Val Cys Ser Asp Arg Pro Gly Leu Leu Phe

850 855 860

TCC ATC AGC AGG GGC ATC TAT AGT TGC ATC CAT GGG GTA CAC ATT GAA 306 Ser Ile Ser Arg Gly Ile Tyr Ser Cys Ile His Gly Val His- Ile Glu

865 870 875

GAA AAG AAG AAA TCT CCA GAT TTC AAT CTG ACT GGA TCA CAG AGC AAC 311 Glu Lys Lys Lys Ser Pro Asp Phe Asn Leu Thr Gly Ser Gin Ser Asn

880 885 890

ATG CTA AAG CTT CTT CGG TCA GCT AAA AAC ATC TCC AAT ATG TCC AAC 315 Met Leu Lys Leu Leu Arg Ser Ala Lys Asn Ile Ser Asn Met Ser Asn

895 900 905

ATG AAC TCC TCA AGA ATG GAC TCA CCT AAA AGA GCT ACT GAC TTC ATT 32 Met Asn Ser Ser Arg Met Asp Ser Pro Lys Arg Ala Thr Asp Phe Ile 910 915 920 925

CAA AGA GGG TCA CTT ATT GTG GAC ATG GTT TCA GAC AAG GGA AAT TTG 32 Gin Arg Gly Ser Leu Ile Val Asp Met Val Ser Asp Lys Gly Asn Leu

930 935 940

ATA TAT TCA GAC AAC AGA TCC TTT CAA GGG AAG GAC AGT ATA TTT GGA 33 Ile Tyr Ser Asp Asn Arg Ser Phe Gin Gly Lys Asp Ser Ile Phe Gly

945 950 955

GAC AAC ATG AAT GAA CTC CAA ACA TTT GTG GCC AAC AGG CAC AAG GAT 33 Asp Asn Met Asn Glu Leu Gin Thr Phe Val Ala Asn Arg His Lys Asp 960 965 970 AAT CTC AGT AAC TAT GTG TTT CAA GGA CAG CAT CCT CTC ACT CTC AAT 3398 Asn Leu Ser Asn Tyr Val Phe Gin Gly Gin His Pro Leu Thr Leu Asn

975 980 985

GAG TCC AAC CCT AAC ACA GTA GAG GTG GCT GTC AGC ACT GAA TCC AAA 3 46 Glu Ser Asn Pro Asn Thr Val Glu Val Ala Val Ser Thr Glu Ser Lys 990 995 1000 1005

GGG AAC TCC CGA CCC CGG CAG CTT TGG AAG AAA TCC ATG GAG TCT CTA 3494 Gly Asn Ser Arg Pro Arg Gin Leu Trp Lys Lys Ser Met Glu Ser Leu

1010 1015 1020

CGC CAG GAT TCT CTA AAC CAG AAC CCA GTC TCC CAG AGG GAT GAG AAG 3542 Arg Gin Asp Ser Leu Asn Gin Asn Pro Val Ser Gin Arg Asp Glu Lys

1025 1030 1035

ACT GCA GAG AAT CGG ACC CAC TCG CTA AAG AGT CCT AGG TAT CTT CCA 3590 Thr Ala Glu Asn Arg Thr His Ser Leu Lys Ser Pro Arg T r Leu Pro

1040 1045 1050

GAA GAG GTA GCC CAC TCT GAC ATT TCA GAA ACC TCA AGC CGG GCC ACA 3638 Glu Glu Val Ala His Ser Asp Ile Ser Glu Thr Ser Ser Arg Ala Thr

1055 1060 1065

TGC CAC AGG GAG CCA GAT AAC AAT AAG AAC CAC AAG ACC AAG GAT AAC 3686 Cys His Arg Glu Pro Asp Asn Asn Lys Asn His Lys Thr Lys Asp Asn 1070 1075 1080 1085

TTC AAA CGG TCA ATG GCC TCT AAG TAT CCC AAG GAC TGT AGC GAT GTT 3734 Phe Lys Arg Ser Met Ala Ser Lys Tyr Pro Lys Asp Cys Ser Asp Val

1090 1095 1100

GAC CGC ACC TAC ATG AAA ACC AAA GCA AGT TCT CCC AGG GAT AAG ATC 3782 Asp Arg Thr Tyr Met Lys Thr Lys Ala Ser Ser Pro Arg Asp Lys Ile

1105 1110 1115

TAT ACC ATT GAT GGT GAG AAG GAG CCC AGC TTC CAC TTA GAT CCT CCT 3830 Tyr Thr Ile Asp Gly Glu Lys Glu Pro Ser Phe His Leu Asp Pro Pro

1120 1125 1130

CAG TTT GTT GAG AAT ATA ACC CTG CCT GAG AAT GTG GGC TTC CCA GAT 3878 Gin Phe Val Glu Asn Ile Thr Leu Pro Glu Asn Val Gly Phe Pro Asp

1135 1140 1145

ACC TAC CAA GAT CAC AAT GAG AAC TTC CGC AAG GGG GAC TCC ACA CTG 392 Thr Tyr Gin Asp His Asn Glu Asn Phe Arg Lys Gly Asp Ser Thr Leu 1150 1155 1160 1165

CCC ATG AAC AGG AAC CCA TTA CAT AAT GAA GAC GGG CTT CCC AAC AAT 397 Pro Met Asn Arg Asn Pro Leu His Asn Glu Asp Gly Leu Pro Asn Asn

1170 1175 1180

GAC CAA TAT AAA CTC TAT GCC AAG CAC TTT ACC TTG AAA GAC AAG GGT 402 Asp Gin Tyr Lys Leu Tyr Ala Lys His Phe Thr Leu Lys Asp Lys Gly

1185 1190 1195

TCC CCA CAC AGT GAG GGC AGT GAT CGA TAC CGG CAG AAC TCC ACA CAT 407 Ser Pro His Ser Glu Gly Ser Asp Arg Tyr Arg Gin Asn Ser Thr His 1200 1205 1210 TGC AGA AGC TGC CTT TCG AAT CTG CCC ACC TAC TCA GGC CAC TTT ACC 41 Cys Arg Ser Cys Leu Ser Asn Leu Pro Thr Tyr Ser Gly His Phe Thr

1215 1220 1225

ATG AGG TCT CCT TTC AAG TGT GAT GCC TGT CTG CGG ATG GGG AAT CTC 41 Met Arg Ser Pro Phe Lys Cys Asp Ala Cys Leu Arg Met Gly Asn Leu 1230 1235 1240 1245

TAT GAC ATT GAT GAA GAC CAG ATG CTT CAG GAG ACA GGT AAC CCA GCT 42 Tyr Asp Ile Asp Glu Asp Gin Met Leu Gin Glu Thr Gly Asn Pro Ala

1250 1255 1260

ACT CGG GAG GAG GTC TAC CAG CAG GAC TGG TCA CAG AAC AAC GCC CTC 42 Thr Arg Glu Glu Val Tyr Gin ^' Gin Asp Trp Ser Gin Asn Asn Ala Leu

1265 1270 1275

CAG TTC CAG AAG AAC AAG CTA AGG ATT AAC CGA CAG CAC TCC TAT GAT 43 Gin Phe Gin Lys Asn Lys Leu Arg Ile Asn Arg Gin His Ser Tyr Asp

1280 1285 1290

AAC ATT CTG GAC AAA CCC AGA GAG ATA GAC CTT AGC AGG CCC TCC CGG 43 Asn Ile Leu Asp Lys Pro Arg Glu Ile Asp Leu Ser Arg Pro Ser Arg

1295 1300 1305

AGC ATA AGC CTC AAG GAC AGG GAA CGG CTA CTG GAG GGC AAC TTG TAT 44 Ser Ile Ser Leu Lys Asp Arg Glu Arg Leu Leu Glu Gly Asn Leu Tyr 1310 1315 1320 1325

GGG AGC CTG TTC AGT GTC CCC TCA AGC AAA CTC TTG GGG AAC AAA AGC 44 Gly Ser Leu Phe Ser Val Pro Ser Ser Lys Leu Leu Gly Asn Lys Ser

1330 1335 1340

TCC CTT TTC CCC CAA GGT CTG GAG GAC AGC AAG AGG AGC AAG TCT CTC 45 Ser Leu Phe Pro Gin Gly Leu Glu Asp Ser Lys Arg Ser Lys Ser Leu

1345 1350 1355

TTG CCA GAC CAC GCC TCT GAT AAT CCT TTC CTC CAC ACG TAT GGG GAT 45 Leu Pro Asp His Ala Ser Asp Asn Pro Phe Leu His Thr Tyr Gly Asp

1360 1365 1370

GAC CAA CGC TTA GTT ATC GGG AGA TGT CCC TCG GAC CCT TAC AAA CAC 45 Asp Gin Arg Leu Val Ile Gly Arg Cys Pro Ser Asp Pro Tyr Lys His

1375 1380 1385

TCA TTG CCA TCA CAG GCG GTA AAT GAC AGC TAT CTT CGG TCA TCC TTG 46 Ser Leu Pro Ser Gin Ala Val Asn Asp Ser Tyr Leu Arg Ser Ser Leu 1390 1395 1400 1405

AGG TCA ACA GCA TCA TAT TGC TCC AGG GAC AGT CGG GGC CAC AGT GAT 46 Arg Ser Thr Ala Ser Tyr Cys Ser Arg Asp Ser Arg Gly His Ser Asp

1410 1415 1420

GTG TAT ATT TCA GAG CAT GTT ATG CCT TAT GCT GCA AAT AAG AAT ACC 47 Val Tyr Ile Ser Glu His Val Met Pro Tyr Ala Ala Asn Lys Asn Thr

1425 1430 1435

ATG TAC TCT ACC CCC AGG GTT TTA AAT TCC TGC AGC AAT AGA CGA GTG 47 Met Tyr Ser Thr Pro Arg Val Leu Asn Ser Cys Ser Asn Arg Arg Val 1440 1445 1450 TAC AAG AAA ATG CCT AGT ATC GAA TCT GAT GTT TAAGATCTTC CATCAGTATT 4843 Tyr Lys Lys Met Pro Ser Ile Glu Ser Asp Val

1455 1460 146

TATCTATAAG GAAACATATA GAATGGCCAA CATTATAGAG GGTAAATGTT GGATGTCCGA 4903 TAGCACCCTA CTAGGAGGAA GAGGGTACAG GGAGGTACTT TTTGTTGGCT CTTTTGCACA 4963 TGGCTCCATG CCATAATCTT CCACTCAAGG AATCTTGTGA GATATGTGCT GAGCACAGCA 5023 TATACCACGT AGGTGAATCC TTAACCAAAA ACAAATAAAT ACACATGGGC AAGTCTCCCA 5083 GACATGGCGA CTGGGCACGG CGGCAATAAT GGTGCATCAG ACGGCGATGG TGACATTGTG 5143 GTT ^• 5146

(2) INFORMATION ABOUT SEQ ID NO: 2:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1464 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: Protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2: Met Gly Arg Leu Gly Tyr Trp Thr Leu Leu Val Leu Pro Ala Leu Leu

1 5 10 15

Val Trp Arg Asp Pro Ala Gin Asn Ala Ala Ala Glu Lys Gly Pro Pro

20 25 30

Ala Leu Asn Ile Ala Val Leu Leu Gly His Ser His Asp Val Thr Glu

35 40 45

Arg Glu Leu Arg Asn Leu Trp Gly Pro Glu Gin Ala Thr Gly Leu Pro

50 55 60

Leu Asp Val Asn Val Val Ala Leu Leu Met Asn Arg Thr Asp Pro Lys 65 70 75 80

Ser Leu Ile Thr His Val Cys Asp Leu Met Ser Gly Ala Arg Ile His

85 90 95

Gly Leu Val Phe Gly Asp Asp Thr Asp Gin Glu Ala Val Ala Gin Met

100 105 110

Leu Asp Phe Ile Ser Ser Gin Thr Phe Ile Pro Ile Leu Gly Ile His

115 120 125

Gly Gly Ala Ser Met Ile Met Ala Asp Lys Asp Pro Thr Ser Thr Phe

130 135 140

Phe Gin Phe Gly Ala Ser Ile Gin Gin Gin Ala Thr Val Met Leu Lys 145 150 155 160

Ile Met Gin Asp Tyr Asp Trp His Val Phe Ser Leu Val Thr Thr Ile

165 170 175

Phe Pro Gly Tyr Arg Asp Phe Ile Ser Phe Ile Lys Thr Thr Val Asp

180 185 190

Asn Ser Phe Val Gly Trp Asp Met Gin Asn Val Ile Thr Leu Asp Thr

195 200 205

Ser Phe Glu Asp Ala Lys Thr Gin Val Gin Leu Lys Lys Ile His Ser

210 215 220

Ser Val Ile Leu Leu Tyr Cys Ser Lys Asp Glu Ala Val Leu Ile Leu 225 230 235 240 Ser Glu Ala Arg Ser Leu Gly Leu Thr Gly Tyr Asp Phe Phe Trp Ile

245 250 255

Val Pro Ser Leu Val Ser Gly Asn Thr Glu Leu Ile Pro Lys Glu Phe

260 265 270

Pro Ser Gly Leu Ile Ser Val Ser Tyr Asp Asp Trp Asp Tyr Ser Leu

275 280 285

Glu Ala Arg Val Arg Asp Gly Leu Gly Ile Leu Thr Thr Ala Ala Ser

290 295 300

Ser Met Leu Glu Lys Phe Ser Tyr Ile Pro Glu Ala Lys Ala Ser Cys 305 310 315 320

Tyr Gly Gin Ala Glu Lys Pro Glu Thr Pro Leu His Thr Leu His Gin

325 330 335

Phe Met Val Asn Val Thr Trp Asp Gly Lys Asp Leu Ser Phe Thr Glu

340 345 350

Glu Gly Tyr Gin Val His Pro Arg Leu Val Val Ile Val Leu Asn Lys

355 360 365

Asp Arg Glu Trp Glu Lys Val Gly Lys Trp Glu Asn Gin Thr Leu Ser

370 375 380

Leu Arg His Ala Val Trp Pro Arg Tyr Lys Ser Phe Ser Asp Cys Glu 385 390 395 400

Pro Asp Asp Asn His Leu Ser Ile Val Thr Leu Glu Glu Ala Pro Phe

405 410 415

Val Ile Val Glu Asp Ile Asp Pro Leu Thr Glu Thr Cys Val Arg Asn

420 425 430

Thr Val Pro Cys Arg Lys Phe Val Lys Ile Asn Asn Ser Thr Asn Glu

435 440 445

Gly Met Asn Val Lys Lys Cys Cys Lys Gly Phe Cys Ile Asp Ile Leu

450 455 460

Lys Lys Leu Ser Arg Thr Val Lys Phe Thr Tyr Asp Leu Tyr Leu Val 465 - 470 475 480

Thr Asn Gly Lys His Gly Lys Lys Val Asn Asn Val Trp Asn Gly Met

485 490 495

Ile Gly Glu Val Val Tyr Gin Arg Ala Val Met Ala Val Gly Ser Leu

500 505 510

Thr Ile Asn Glu Glu Arg Ser Glu Val Val Asp Phe Ser Val Pro Phe

515 520 525

Val Glu Thr Gly Ile Ser Val Met Val Ser Arg Ser Asn Gly Thr Val

530 535 540

Ser Pro Ser Ala Phe Leu Glu Pro Phe Ser Ala Ser Val Trp Val Met 545 550 555 560

Met Phe Val Met Leu Leu Ile Val Ser Ala Ile Ala Val Phe Val Phe

565 570 575

Glu Tyr Phe Ser Pro Val Gly Tyr Asn Arg Asn Leu Ala Lys Gly Lys

580 585 590

Ala Pro His Gly Pro Ser Phe Thr Ile Gly Lys Ala Ile Trp Leu Leu 595 600 605 Trp Gly Leu Val Phe Asn Asn Ser Val Pro Val Gin Asn Pro Lys Gly

610 - 615 620

Thr Thr Ser Lys Ile Met Val Ser Val Trp Ala Phe Phe Ala Val Ile 625 630 635 640

Phe Leu Ala Ser Tyr Thr Ala Asn Leu Ala Ala Phe Met Ile Gin Glu

645 650 655

Glu Phe Val Asp Gin Val Thr Gly Leu Ser Asp Lys Lys Phe Gin Arg

660 665 670

Pro His Asp Tyr Ser Pro Pro Phe Arg Phe Gly Thr Val Pro Asn Gly

675 680 685

Ser Thr Glu Arg Asn Ile Arg Äsn Asn Tyr Pro Tyr Met His Gin Tyr

690 695 700

Met Thr Arg Phe Asn Gin Arg Gly Val Glu Asp Ala Leu Val Ser Leu 705 710 715 720

Lys Thr Gly Lys Leu Asp Ala Phe Ile Tyr Asp Ala Ala Val Leu Asn

725 730 735

Tyr Lys Ala Gly Arg Asp Glu Gly Cys Lys Leu Val Thr Ile Gly Ser

740 745 750

Gly Tyr Ile Phe Ala Ser Thr Gly Tyr Gly Ile Ala Leu Gin Lys Gly

755 760 765

Ser Pro Trp Lys Arg Gin Ile Asp Leu Ala Leu Leu Gin Phe Val Gly

770 775 780

Asp Gly Glu Met Glu Glu Leu Glu Thr Leu Trp Leu Thr Gly Ile Cys 785 790 795 800

His Asn Glu Lys Asn Glu Val Met Ser Ser Gin Leu Asp Ile Asp Asn

805 810 815

Met Ala Gly Val Phe Tyr Met Leu Ala Ala Ala Met Ala Leu Ser Leu

820 825 830

Ile Thr Phe Ile Trp Glu His Leu Phe Tyr Trp Lys Leu Arg Phe Cys

835 840 845

Phe Thr Gly Val Cys Ser Asp Arg Pro Gly Leu Leu Phe Ser Ile Ser

850 855 860

Arg Gly Ile Tyr Ser Cys Ile His Gly Val His Ile Glu Glu Lys Lys 865 870 875 880

Lys Ser Pro Asp Phe Asn Leu Thr Gly Ser Gin Ser Asn Met Leu Lys

885 890 895

Leu Leu Arg Ser Ala Lys Asn Ile Ser Asn Met Ser Asn Met Asn Ser

900 905 910

Ser Arg Met Asp Ser Pro Lys Arg Ala Thr Asp Phe Ile Gin Arg Gly

915 920 925

Ser Leu Ile Val Asp Met Val Ser Asp Lys Gly Asn Leu Ile Tyr Ser

930 935 940

Asp Asn Arg Ser Phe Gin Gly Lys Asp Ser Ile Phe Gly Asp Asn Met 945 950. 955 960

Asn Glu Leu Gin Thr Phe Val Ala Asn Arg His Lys Asp Asn Leu Ser 965 970 975 Asn Tyr Val Phe Gin Gly Gin His Pro Leu Thr Leu Asn Glu Ser Asn

980 985 990

Pro Asn Thr Val Glu Val Ala Val Ser Thr Glu Ser Lys Gly Asn Ser

995 1000 1005

Arg Pro Arg Gin Leu Trp Lys Lys Ser Met Glu Ser Leu Arg Gin Asp

1010 1015 1020

Ser Leu Asn Gin Asn Pro Val Ser Gin Arg Asp Glu Lys Thr Ala Glu 1025 1030 1035 1040

Asn Arg Thr His Ser Leu Lys Ser Pro Arg Tyr Leu Pro Glu Glu Val

1045 1050 1055

Ala His Ser Asp Ile Ser Glu Thr Ser Ser Arg Ala Thr Cys His Arg

1060 1065 1070

Glu Pro Asp Asn Asn Lys Asn His Lys Thr Lys Asp Asn Phe Lys Arg

1075 1080 1085

Ser Met Ala Ser Lys Tyr Pro Lys Asp Cys Ser Asp Val Asp Arg Thr

1090 1095 1100

Tyr Met Lys Thr Lys Ala Ser Ser Pro Arg Asp Lys Ile Tyr Thr Ile 1105 1110 1115 1120

Asp Gly Glu Lys Glu Pro Ser Phe His Leu Asp Pro Pro Gin Phe Val

1125 1130 1135

Glu Asn Ile Thr Leu Pro Glu Asn Val Gly Phe Pro Asp Thr Tyr Gin

1140 1145 1150

Asp His Asn Glu Asn Phe Arg Lys Gly Asp Ser Thr Leu Pro Met Asn

1155 1160 1165

Arg Asn Pro Leu His Asn Glu Asp Gly Leu Pro Asn Asn Asp Gin Tyr

1170 1175 1180

Lys Leu Tyr Ala Lys His Phe Thr Leu Lys Asp Lys Gly Ser Pro His 1185 1190 1195 1200

Ser Glu Gly Ser Asp Arg Tyr Arg Gin Asn Ser Thr His Cys Arg Ser

1205 1210 1215

Cys Leu Ser Asn Leu Pro Thr Tyr Ser Gly His Phe Thr Met Arg Ser

1220 1225 1230

Pro Phe Lys Cys Asp Ala Cys Leu Arg Met Gly Asn Leu Tyr Asp Ile

1235 1240 1245

Asp Glu Asp Gin Met Leu Gin Glu Thr Gly Asn Pro Ala Thr Arg Glu

1250 1255 1260

Glu Val Tyr Gin Gin Asp Trp Ser Gin Asn Asn Ala Leu Gin Phe Gin 1265 1270 1275 1280

Lys Asn Lys Leu Arg Ile Asn Arg Gin His Ser Tyr Asp Asn Ile Leu

1285 1290 1295

Asp Lys Pro Arg Glu Ile Asp Leu Ser Arg Pro Ser Arg Ser Ile Ser

1300 1305 1310

Leu Lys Asp Arg Glu Arg Leu Leu Glu Gly Asn Leu Tyr Gly Ser Leu

1315 1320 1325

Phe Ser Val Pro Ser Ser Lys Leu Leu Gly Asn Lys Ser Ser Leu Phe 1330 1335 1340 Pro Gin Gly Leu Glu Asp Ser Lys Arg Ser Lys Ser Leu Leu Pro Asp 1345 1350 1355 1360

His Ala Ser Asp Asn Pro Phe Leu His Thr Tyr Gly Asp Asp Gin Arg

1365 1370 1375

Leu Val Ile Gly Arg Cys Pro Ser Asp Pro Tyr Lys His Ser Leu Pro

1380 1385 1390

Ser Gin Ala Val Asn Asp Ser Ty Leu Arg Ser Ser Leu Arg Ser Thr

1395 1400 1405

Ala Ser Tyr Cys Ser Arg Asp Ser Arg Gly His -Ser Asp Val Tyr Ile

1410 1415 1420

Ser Glu His Val Met Pro Tyr Ala Ala Asn Lys Asn Thr Met Tyr Ser 1425 1430 1435 1440

Thr Pro Arg Val Leu Asn Ser Cys Ser Asn Arg Arg Val Tyr Lys Lys

1445 1450 1455

Met Pro Ser Ile Glu Ser Asp Val

1460 (2) INFORMATION ON SEQ ID NO: 3: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 5359 base pairs

(B) TYPE: nucleic acid

(C) STRAND FORM; singles

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

(iii) HYPOTHETICAL: NO (iii) ANTISENSE: NO (vi) ORIGINAL ORIGIN:

(A) ORGANISM: Rattus norvegicus .- (F) FABRIC TYPE: Brain (ix) CHARACTERISTICS:

(A) NAME / KEY: CDS

(B) LOCATION: 857..5305

(Xi) SEQÜENZBESCHREIBÜNG: SEQ ID NO: 3: TCCCCCGGCT CCCATCACCA CCAACTGGAA TCTGGGCTCT CCTGCCTCTC CACCCCCCCC 6 CCTTTTTGAG AAAGGAAGGT CCTCCCCCAC CCCCTTTCCA GCACCACTTC CTTGGGTTCC 12 TTCACCTCTA GGAAATCTCG GGGCTTGGCT CAATGGAGAA GCGCTTCTCG GGTCAAGCTG 18 CCTCTCCATC CCCGGCATCC AGCGGAGGCT CATCCGAAAT ACATTTCCTT CTGCTCTGCA 24 AAGAGTCGTA GTGAACTTCC CAACCCGAGA GACTGATCAT CTATCCTGCT TTGCAGAAAG 30 GAAATCTCTC AAACCCTCGA CACCCGCCAC CCCTCTCCGA AGAGTGCCAC CGCGGAGGTG 36 CTCAGAACCG GAAGGGACGC TTTGGGAATG ACCATCGTCT ACGGAGGGAC AGAGCCTGCC 42 CCCAGCTTCT CCACACAGAG ACTCCTCTAC TAAGCTCCAA AAÄTTAAAAG AAAAAAGAAA 48 AAGAAAAAAG ATCAATCTGT AATTGCTGAA GACTCCTTAA ATATATATAT ATATATATAT 54 ATTCGGGCTA CTAACCTCAC ATGCACATGG GATAATGACT CTGGATTCTG CATTGTGAGC 60 TGCTCTCCAC ACCCTGAGAT CCCCTCTTAC ATTACATTTT TTCCTTTGAA TTTGCATCTC 6 GTCAAGACAC AAGATTAAAA CCGAAATTTA CACTACACTG GATTTTAAAT TTTCTTCCGT 72 TCCTTTATCC TCCGTCTTTC TTATGTGGAT ATGCAAGCGA GAAGAGGAGC CCTGGATATT 7 CCCAACATGC TCTCTCCCTT AATCTGTCCG CCTAGAGGTT TGGCGTCTAC AAACCAAG AG 8 AGCCGACTAG CTGAAG ATG AAG CCC AGC GCA GAG TGC TGT TCC CCC AAG 88

Met Lys Pro Ser Ala Glu Cys Cys Ser Pro Lys 1 5 10

TTC TGG TTG GTG TTG GCC GTC TTG GCC GTA TCA GGC AGC AAA GCT CGT 93 Phe Trp Leu Val Leu Ala Val Leu Ala Val Ser Gly Ser Lys Ala Arg

15 20 25

TCC CAA AAG AGC CCC CCC AGC ATC GGC ATC GCT GTC ATC CTC GTG GGC 98 Ser Gin Lys Ser Pro Pro Ser Ile Gly Ile Ala Val Ile Leu Val Gly

30 35 40

ACT TCA GAC GAA GTG GCC ATA AAA GAC GCC CAC GAG AAA GAT GAC TTC 103 Thr Ser Asp Glu Val Ala Ile Lys Asp Ala His Glu Lys Asp Asp Phe

45 50 55

CAT CAT CTC TCA GTA GTT CCC CGG GTG GAG CTG GTA GCC ATG AAC GAA 108 His His Leu Ser Val Val Pro Arg Val Glu Leu Val Ala Met Asn Glu 60 65 70 75

ACT GAC CCA AAG AGC ATC ATC ACC CGT ATC TGC GAT CTT ATG TCT GAC 112 Thr Asp Pro Lys Ser Ile Ile Thr Arg Ile Cys Asp Leu Met Ser Asp

80 85 90

CGG AAG ATC CAG GGG GTG GTG TTC GCG GAT GAC ACC GAC CAA GAA GCC 117 Arg Lys Ile Gin Gly Val Val Phe Ala Asp Asp Thr Asp Gin Glu Ala

95 100 105

ATC GCT CAG ATC CTC GAC TTC ATT TCT GCT CAG ACT CTC ACC CCC ATC 122 Ile Ala Gin Ile Leu Asp Phe Ile Ser Ala Gin Thr Leu Thr Pro Ile

110 115 120

CTG GGC ATC CAT GGG GGC TCA TCT ATG ATA ATG GCG GAT AAG GAT GAG 127 Leu Gly Ile His Gly Gly Ser Ser Met Ile Met Ala Asp Lys Asp Glu

125 130 135

TCC TCC ATG TTC TTC CAG TTT GGC CCG TCT ATC GAA CAG CAA GCT TCC 132 Ser Ser Met Phe Phe Gin Phe Gly Pro Ser Ile Glu Gin Gin Ala Ser 140 145 150 155

GTC ATG CTC AAC ATC ATG GAA GAA TAT GAC TGG TAC ATC TTT TCC ATC 13 Val Met Leu Asn Ile Met Glu Glu Tyr Asp Trp Tyr Ile Phe Ser Ile

160 165 170

GTC ACC ACC TAC TTC CCT GGC TAC CAG-GAC TTT GTG AAC AAG ATC CGC 14 Val Thr Thr Tyr Phe Pro Gly Tyr Gin Asp Phe Val Asn Lys Ile Arg

175 180 185

AGT ACC ATC GAG AAC AGC TTC GTG GGC TGG GAG CTC GAG GAA GTC CTC 14 Ser Thr Ile Glu Asn Ser Phe Val Gly Trp Glu Leu Glu Glu Val Leu

190 195 200

CTG CTA GAC ATG TCT CTG GAC GAT GGC GAC TCT AAG ATT CAG AAT CAG 15 Leu Leu Asp Met Ser Leu Asp Asp Gly Asp Ser Lys Ile Gin Asn Gin

205 210 215

CTG AAG AAG CTC CAA AGC CCC ATC ATT CTC CTT TAT TGC ACG AAG GAG 15 Leu Lys Lys Leu Gin Ser Pro Ile Ile Leu Leu Tyr Cys Thr Lys Glu 220 225 230 235 GAA GCC ACC TAC ATT TTT GAA GTA GCT AAC TCA GTT GGG CTG ACT GGC 1609 Glu Ala Thr Tyr Ile Phe Glu Val Ala Asn Ser Val Gly Leu Thr Gly

240 245 250

TAC GGC TAC ACG TGG ATT GTG CCG AGT CTG GTG GCC GGG GAT ACG GAC 1657 Tyr Gly Tyr Thr Trp Ile Val Pro Ser Leu Val Ala Gly Asp Thr Asp

255 260 265

ACG GTG CCT TCA GAG TTC CCC ACG GGG CTT ATC TCT GTG TCT TAT GAT 1705 Thr Val Pro Ser Glu Phe Pro Thr Gly Leu Ile Ser Val Ser Tyr Asp

270 275 ^• 280

GAA TGG GAC TAT GGC CTT CCT GCC AGA GTG AGA GAT GGA ATT GCC ATC 1753 Glu Trp Asp Tyr Gly Leu Pro -Ala Arg Val Arg Asp Gly Ile Ala Ile

285 290 295

ATC ACC ACT GCT GCC TCG GAC ATG CTG TCC GAA CAC AGT TTC ATC CCT 1801 Ile Thr Thr Ala Ala Ser Asp Met Leu Ser Glu His Ser Phe Ile Pro 300 305 310 315

GAG CCC AAG AGC AGT TGC TAC AAC ACC CAC GAG AAG AGG ATC TAC CAG 1849 Glu Pro Lys Ser Ser Cys Tyr Asn Thr His Glu Lys Arg Ile Tyr Gin

320 325 330

TCT AAC ATG TTG AAT AGG TAT CTG ATC AAT GTC ACT TTT GAA GGG AGA 1897 Ser Asn Met Leu Asn Arg Tyr Leu Ile Asn Val Thr Phe Glu Gly Arg

335 340 345

AAC CTG TCC TTC AGC GAA GAT GGC TAC CAG ATG CAT CCG AAG CTG GTG 1945 Asn Leu Ser Phe Ser Glu Asp Gly Tyr Gin Met His Pro Lys Leu Val

350 355 360

ATA ATC CTT CTG AAC AAG GAG AGG AAG TGG GAG AGG GTG GGG AAA TGG 1993 Ile Ile Leu Leu Asn Lys Glu Arg Lys Trp Glu Arg Val Gly Lys Trp

365 370 375

AAG GAC AAG TCC CTG CAG ATG AAG TAT TAT GTG TGG CCT CGG ATG TGT 204 Lys Asp Lys Ser Leu Gin Met Lys Tyr Tyr Val Trp Pro Arg Met Cys 380 385 - 390 395

CCT GAG ACT GAG GAG CAA GAG GAT GAC CAT CTG AGC ATT GTC ACC TTG 208 Pro Glu Thr Glu Glu Gin Glu Asp Asp His Leu Ser Ile Val Thr Leu

400 405 410

GAG GAG GCG CCA TTT GTC ATT GTG GAA AGC GTG GAC CCT CTC AGT GGA 213 Glu Glu Ala Pro Phe Val Ile Val Glu Ser Val Asp Pro Leu Ser Gly

415 420 425

ACC TGC ATG AGG AAT ACA GTC CCG TGC CAG AAG CGC ATC ATC TCT GAG 218 Thr Cys Met Arg Asn Thr Val Pro Cys Gin Lys Arg Ile Ile Ser Glu

430 435 440

AAT AAA ACA GAT GAG GAA CCA GGC TAC ATC AAA AAA TGC TGC AAG GGG 223 Asn Lys Thr Asp Glu Glu Pro Gly Tyr Ile Lys Lys Cys Cys Lys Gly

445 450 455

TTC TGT ATT GAC ATC CTT AAG AAA ATT TCT AAG TCT GTG AAG TTC ACC 228 Phe Cys Ile Asp Ile Leu Lys Lys Ile Ser Lys Ser Val Lys Phe Thr 460 465 470 475 TAT GAC CTT TAC CTG GTG ACC AAT GGC AAG CAC GGG AAG AAG ATT AAT 232 Tyr Asp Leu Tyr Leu Val Thr Asn Gly Lys His Gly Lys Lys Ile Asn

480 485 490

GGG ACC TGG AAT GGC ATG ATC GGT GAG GTG GTC ATG AAG AGG GCC TAC 237 Gly Thr Trp Asn Gly Met Ile Gly Glu Val Val Met Lys Arg Ala Tyr

495 500 505

ATG GCA GTG GGA TCA CTA ACT ATC AAT GAA GAA CGG TCA GAG GTG GTT 242 Met Ala Val Gly Ser Leu Thr Ile Asn Glu Glu Arg Ser Glu Val Val

510 515 520

GAC TTC TCT GTA CCC TTC ATA GAA ACT GGC ATC AGT GTC ATG GTA TCT 247 Asp Phe Ser Val Pro Phe Ile Glu Thr Gly Ile Ser Val Met Val Ser

525 530 535

CGC AGC AAT GGG ACT GTG TCA CCT TCT GCC TTC TTA GAG CCA TTC AGC 252 Arg Ser Asn Gly Thr Val Ser Pro Ser Ala Phe Leu Glu Pro Phe Ser 540 545 550 555

GCT GAC GTG TGG GTG ATG ATG TTT GTG ATG CTG CTC ATT GTT TCT GCG 256 Ala Asp Val Trp Val Met Met Phe Val Met Leu Leu Ile Val Ser Ala

560 565 570

GTG GCT GTC TTT GTC TTT GAA TAC TTC AGC CCT GTG GGT TAC AAC AGG 261 Val Ala Val Phe Val Phe Glu Tyr Phe Ser Pro Val Gly Tyr Asn Arg

575 580 585

TGC CTA GCC GAT GGC AGA GAG CCA GGA GGC CCA TCT TTC ACC ATC GGC 266 Cys Leu Ala Asp Gly Arg Glu Pro Gly Gly Pro Ser Phe Thr Ile Gly

590 595 600

AAA GCA ATT TGG TTA CTC TGG GGT CTG GTG TTT AAC AAC TCC GTA CCT 271 Lys Ala Ile Trp Leu Leu Trp Gly Leu Val Phe Asn Asn Ser Val Pro

605 610 615

GTG CAG AAC CCA AAG GGG ACC ACC TCC AAG ATC ATG GTG TCA GTG TGG 276 Val Gin Asn Pro Lys Gly Thr Thr Ser Lys Ile Met Val Ser Val Trp 620 625 630 635

GCC TTC TTT GCT GTC ATT TTC CTG GCC AGC TAC ACT GCC AAC TTA GCA 280 Ala Phe Phe Ala Val Ile Phe Leu Ala Ser Tyr Thr Ala Asn Leu Ala

640 645 650

GCC TTC ATG ATC CAA GAG GAG TAT GTG GAC CAG GTT TCT GGC CTG AGT 285 Ala Phe Met Ile Gin Glu Glu Tyr Val Asp Gin Val Ser Gly Leu Ser

655 660 665

GAC AAG AAG TTC CAG AGA CCT AAT GAC TTC TCA CCC CCT TTC CGC TTT 290 Asp Lys Lys Phe Gin Arg Pro Asn Asp Phe Ser Pro Pro Phe Arg Phe

670 675 680

GGG ACT GTG CCC AAT GGC AGC ACA GAG AGG AAT ATC CGT AAT AAC TAT 295 Gly Thr Val Pro Asn Gly Ser Thr Glu Arg Asn Ile Arg Asn Asn Tyr

685 690 695

GCA GAA ATG CAT GCC TAC ATG GGA AAG TTC AAC CAA AGG GGT GTA GAT 300 Ala Glu Met His Ala Tyr Met Gly Lys Phe Asn Gin Arg Gly Val Asp 700 705 710 715 GAT GCA TTG CTC TCC CTG AAA ACA GGG AAG CTT GAT GCA TTC ATC TAT 3049 Asp Ala Leu Leu Ser Leu Lys Thr Gly Lys Leu Asp Ala Phe Ile Tyr

720 725 730

GAT GCA GCT GTG CTC AAC TAC ATG GCT GGA AGG GAC GAA GGC TGC AAA 3097 Asp Ala Ala Val Leu Asn Tyr Met Ala Gly Arg Asp Glu Gly Cys Lys

735 740 745

CTG GTG ACC ATT GGC AGT GGC AAG GTC TTT GCT TCT ACC GGC TAT GGC 3145 Leu Val Thr Ile Gly Ser Gly Lys Val Phe Ala Ser Thr Gly Tyr Gly

750 755 760

ATT GCT ATC CAA AAG GAC TCC GGG TGG AAG CGC CAG GTG GAC CTG GCT 3193 Ile Ala Ile Gin Lys Asp Ser Gly Trp Lys Arg Gin Val Asp Leu Ala

765 770 775

ATC CTG CAG CTG TTT GGA GAT GGG GAG ATG GAA GAA CTG GAA GCT CTC 3241 Ile Leu Gin Leu Phe Gly Asp Gly Glu Met Glu Glu Leu Glu Ala Leu 780 785 790 795

TGG CTC ACT GGC ATT TGC CAC AAT GAG AAG AAT GAG GTG ATG AGC AGC 3289 Trp Leu Thr Gly Ile Cys His Asn Glu Lys Asn Glu Val Met Ser Ser

800 805 810

CAG CTG GAC ATC GAC AAT ATG GCA GGT GTC TTC TAT ATG TTG GGG GCA 3337 Gin Leu Asp Ile Asp Asn Met Ala Gly Val Phe Tyr Met Leu Gly Ala

815 ^" 820 825

GCC ATG GCC CTC AGC CTC ATC ACC TTC ATC TGT GAG CAT CTG TTC TAT 3385 Ala Met Ala Leu Ser Leu Ile Thr Phe Ile Cys Glu His Leu Phe Tyr

830 835 840

TGG CAG TTC CGG CAT TGC TTC ATG GGT GTC TGT TCT GGC AAG CCT GGC 3433 Trp Gin Phe Arg His Cys Phe Met Gly Val Cys Ser Gly Lys Pro Gly

845 850 855

ATG GTC TTC TCC ATC AGC AGA GGT ATC TAC AGC TGT ATC CAT GGG GTA 3481 Met Val Phe Ser Ile Ser Arg Gly Ile Tyr Ser Cys Ile His Gly Val 860 865 870 875

GCC ATA GAG GAG CGC CAA TCC GTG ATG AAC TCC CCC ACT GCC ACC ATG 3529 Ala Ile Glu Glu Arg Gin Ser Val Met Asn Ser Pro Thr Ala Thr Met

880 885 890

AAC AAC ACC CAC TCC AAC ATC CTA CGC TTG CTC CGC ACG GCC AAG AAC 3577 Asn Asn Thr His Ser Asn Ile Leu Arg Leu Leu Arg Thr Ala Lys Asn

895 900 905

ATG GCC AAC CTG TCT GGA GTA AAC GGC TCC CCT CAG AGT GCC CTG GAC 3625 Met Ala Asn Leu Ser Gly Val Asn Gly Ser Pro Gin Ser Ala Leu Asp

910 915 920

TTC ATC CGC CGA GAG TCC TCC GTC TAC GAC ATC TCT GAG CAT CGT CGC 3673 Phe Ile Arg Arg Glu Ser Ser Val Tyr Asp Ile Ser Glu His Arg Arg

925 930 935

AGC TTC ACG CAT TCA GAC TGC AAG TCT TAC AAT AAC CCA CCC TGT GAG 372 Ser Phe Thr His Ser Asp Cys Lys Ser Tyr Asn Asn Pro Pro Cys Glu 940 945 950 955 GAA AAC CTG TTC AGT GAC TAC ATT AGC GAG GTA GAG AGA ACA TTT GGT 376 Glu Asn Leu Phe Ser Asp Tyr Ile Ser Glu Val Glu Arg Thr Phe Gly

960 965 970

AAC CTG CAG CTG AAG GAC AGC AAT GTG TAC CAA GAC CAC TAT CAC CAT 381 Asn Leu Gin Leu Lys Asp Ser Asn Val Tyr Gin Asp His Tyr His His

975 980 985

CAC CAC CGG CCA CAC AGC ATC GGC AGC ACC AGC TCC ATT GAT GGG CTC 386 His His Arg Pro His Ser Ile Gly Ser Thr Ser Ser Ile Asp Gly Leu

990 995 1000

TAT GAC TGT GAC AAC CCA CCC TTC ACC ACC CAG CCC AGG TCA ATC AGC 391 Tyr Asp Cys Asp Asn Pro Pro Phe Thr Thr Gin Pro Arg Ser Ile Ser

1005 1010 1015

AAG AAA CCC CTG GAC ATC GGC CTG CCC TCC TCC AAA CAT AGC CAG CTC 396 Lys Lys Pro Leu Asp Ile Gly Leu Pro Ser Ser Lys His Ser Gin Leu 1020 1025 1030 1035

AGC GAC CTG TAT GGC AAG TTC TCT TTC AAG AGT GAC CGC TAC AGT GGC 400 Ser Asp Leu Tyr Gly Lys Phe Ser Phe Lys Ser Asp Arg Tyr Ser Gly

1040 1045 1050

CAC GAC GAC TTG ATT CGA TCG GAT GTC TCC GAC ATC TCC ACG CAC ACT 405 His Asp Asp Leu Ile Arg Ser Asp Val Ser Asp Ile Ser Thr His Thr

1055 1060 1065

GTC ACC TAT GGG AAC ATC GAG GGC AAC GCA GCC AAG AGG AGG AAA CAG 410 Val Thr T r Gly Asn Ile Glu Gly Asn Ala Ala Lys Arg Arg Lys Gin

1070 1075 1080

CAG TAT AAG GAC AGT CTA AAG AAG CGG CCA GCC TCG GCC AAA TCG AGG 415 Gin Tyr Lys Asp Ser Leu Lys Lys Arg Pro Ala Ser Ala Lys Ser Arg

1085 1090 1095

AGG GAG TTT GAT GAA ATC GAG CTG GCC TAC CGT CGC CGA CCA CCC CGC 420 Arg Glu Phe Asp Glu Ile Glu Leu Ala Tyr Arg Arg Arg Pro Pro Arg 1100 1105 1110 1115

TCC CCG GAC CAC AAG CGC TAC TTC AGG GAC AAA GAA GGG CTC CGA GAC 424 Ser Pro Asp His Lys Arg Tyr Phe Arg Asp Lys Glu Gly Leu Arg Asp

1120 1125 1130

TTC TAC CTG GAC CAG TTC CGA ACA AAG GAG AAC TCG CCT CAC TGG GAG 429 Phe Tyr Leu Asp Gin Phe Arg Thr Lys Glu Asn Ser Pro His Trp Glu

1135 1140 1145

CAC GTG GAC TTG ACT GAC ATT TAC AAA GAA CGC AGT GAC GAC TTC AAG 43 His Val Asp Leu Thr Asp Ile Tyr Lys Glu Arg Ser Asp Asp Phe Lys

1150 1155 1160

CGA GAT TCG GTC AGT GGA GGT GGG CCC TGT ACC AAC AGG TCT CAC CTC 43 Arg Asp Ser Val Ser Gly Gly Gly Pro Cys Thr Asn Arg Ser His Leu

1165 1170 1175

AAA CAC GGA ACG GGC GAG AAG CAC GGA GTG GTA GGC GGG GTG CCT GCT 44 Lys His Gly Thr Gly Glu Lys His Gly Val Val Gly Gly Val Pro Ala 1180 1185 1190 1195 CCT TGG GAG AAG AAC CTG ACC AAT GTG GAT TGG GAG GAC CGG TCT GGG 4489 Pro Trp Glu Lys Asn Leu Thr Asn Val Asp Trp Glu Asp Arg Ser Gly

1200 1205 1210

GGC AAC TTC TGC CGC AGC TGT CCT TCC AAG CTG CAC AAT TAC TCC TCG 4537 Gly Asn Phe Cys Arg Ser Cys Pro Ser Lys Leu His Asn Tyr Ser Ser

1215 1220 1225

ACG GTG GCA GGG CAG AAC TCG GGC CGG CAG GCC TGC ATC AGA TGT GAG 4585 Thr Val Ala Gly Gin Asn Ser Gly Arg Gin Ala Cys Ile Arg Cys Glu

1230 1235 ^• 1240

GCC TGT AAG AAG GCT GGT AAC CTG TAC GAC ATC AGC GAG GAC AAC TCC 4633 Ala Cys Lys Lys Ala Gly Asn Leu Tyr Asp Ile Ser Glu Asp Asn Ser

1245 1250 1255

CTG CAG GAA CTG GAC CAG CCG GCT GCC CCC GTG GCT GTG ACA TCC AAC 4681 Leu Gin Glu Leu Asp Gin Pro Ala Ala Pro Val Ala Val Thr Ser Asn 1260 1265 1270 1275

GCC TCC AGC ACC AAG TAC CCT CAA AGC CCG ACT AAT TCC AAG GCT CAG 4729 Ala Ser Ser Thr Lys Tyr Pro Gin Ser Pro Thr Asn Ser Lys Ala Gin

1280 1285 1290

AAG AAG AAT CGG AAC AAA CTG CGC CGG CAG CAT TCC TAC GAC ACC TTC 4777 Lys Lys Asn Arg Asn Lys Leu Arg Arg Gin His Ser Tyr Asp Thr Phe

1295 1300 1305

GTG GAC CTG CAG AAG GAG GAG GCC GCC TTG GCC CCA CGC AGC GTG AGC 4825 Val Asp Leu Gin Lys Glu Glu Ala Ala Leu Ala Pro Arg Ser Val Ser

1310 1315 1320

CTG AAA GAC AAG GGC CGA TTC ATG GAT GGG AGC CCC TAC GCC CAT ATG 4873 Leu Lys Asp Lys Gly Arg Phe Met Asp Gly Ser Pro Tyr Ala His Met

1325 1330 1335

TTT GAG ATG CCA GCT GGT GAG AGC TCC TTT GCC AAC AAG TCC TCA GTG 4921 Phe Glu Met Pro Ala Gly Glu Ser Ser Phe Ala Asn Lys Ser Ser Val 1340 1345 1350 1355

CCC ACT GCC GGA CAC CAC CAC AAC AAC CCC GGC AGC GGC TAC ATG CTC 4969 Pro Thr Ala Gly His His His Asn Asn Pro Gly Ser Gly Tyr Met Leu

1360 1365 1370

AGC AAG TCG CTC TAC CCT GAC CGG GTC ACG CAA AAC CCT TTC ATC CCC 5017 Ser Lys Ser Leu Tyr Pro Asp Arg Val Thr Gin Asn Pro Phe Ile Pro

1375 1380 1385

ACT TTT GGG GAT GAC CAG TGC TTG CTT CAC GGC AGC AAA TCC TAC TTC 506 Thr Phe Gly Asp Asp Gin Cys Leu Leu His Gly Ser Lys Ser Tyr Phe

1390 1395 1400

TTC AGG CAG CCC ACG GTG GCA GGG GCG TCA AAA ACA AGG CCG GAC TTC 511 Phe Arg Gin Pro Thr Val Ala Gly Ala Ser Lys Thr Arg Pro Asp Phe

1405 1410 ^• 1415

CGG GCC CTT GTC ACC AAT AAG CCA GTG GTG GTC ACC CTT CAT GGG GCT 516 Arg Ala Leu Val Thr Asn Lys Pro Val Val Val Thr Leu His Gly Ala 1420 1425 1430 1435 GTG CCA GGT CGT TTC CAG AAG GAC ATT TGT ATA GGG AAC CAG TCC AAC 520 Val Pro Gly Arg Phe Gin Lys Asp Ile Cys Ile Gly Asn Gin Ser Asn

1440 1445 1450

CCC TGT GTG CCT AAC AAC AAA AAC CCC AGG GCT TTC AAT GGC TCC AGC 525 Pro Cys Val Pro Asn Asn Lys Asn Pro Arg Ala Phe Asn Gly Ser Ser

1455 1460 1465

AAT GGA CAT GTT TAT GAG AAA CTT TCT AGT ATT GAG TCT GAT GTC TGAGTGAGGG Asn Gly His Val Tyr Glu Lys Leu Ser Ser Ile Glu Ser Asp Val

1470 1475 1480

AAGAGAGAGA GGTTAAGGTG GGTACGGGAG GGATAGGGCT GTGGGCC 535

(2) INFORMATION ABOUT SEQ ID NO: 4:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1482 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: Protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4: Met Lys Pro Ser Ala Glu Cys Cys Ser Pro Lys Phe Trp Leu Val Leu

1 5 10 15

Ala Val Leu Ala Val Ser Gly Ser Lys Ala Arg Ser Gin Lys Ser Pro

20 25 30

Pro Ser Ile Gly Ile Ala Val Ile Leu Val Gly Thr Ser Asp Glu Val

35 40 45

Ala Ile Lys Asp Ala His Glu Lys Asp Asp Phe His His Leu Ser Val

50 55 60

Val Pro Arg Val Glu Leu Val Ala Met Asn Glu Thr Asp Pro Lys Ser 65 70 75 80

Ile Ile Thr Arg Ile Cys Asp Leu Met Ser Asp Arg Lys Ile Gin Gly

85 90 95

Val Val Phe Ala Asp Asp Thr Asp Gin Glu Ala Ile Ala Gin Ile Leu

100 105 110

Asp Phe Ile Ser Ala Gin Thr Leu Thr Pro Ile Leu Gly Ile His Gly

115 120 125

Gly Ser Ser Met Ile Met Ala Asp Lys Asp Glu Ser Ser Met Phe Phe

130 135 140

Gin Phe Gly Pro Ser Ile Glu Gin Gin Ala Ser Val Met Leu Asn Ile 145 150 155 160

Met Glu Glu Tyr Asp Trp Tyr Ile Phe Ser Ile Val Thr Thr Tyr Phe

165 170 175

Pro Gly Tyr Gin Asp Phe Val Asn Lys Ile Arg Ser Thr Ile Glu Asn

180 185 190

Ser Phe Val Gly Trp Glu Leu Glu Glu Val Leu Leu Leu Asp Met Ser

195 200 205

Leu Asp Asp Gly Asp Ser Lys Ile Gin Asn Gin Leu Lys Lys Leu Gin

210 215 220

Ser Pro Ile Ile Leu Leu Tyr Cys Thr Lys Glu Glu Ala Thr Tyr Ile 225 230 235 240 Phe Glu Val Ala Asn Ser Val Gly Leu Thr Gly Tyr Gly Tyr Thr Trp

- 245 250 255

Ile Val Pro Ser Leu Val Ala Gly Asp Thr Asp Thr Val Pro Ser Glu

260 265 270

Phe Pro Thr Gly Leu Ile Ser Val Ser Tyr Asp Glu Trp Asp Tyr Gly

275 280 285

Leu Pro Ala Arg Val Arg Asp Gly Ile Ala Ile Ile Thr Thr Ala Ala

290 295 300

Ser Asp Met Leu Ser Glu His Ser Phe Ile Pro Glu Pro Lys Ser Ser 305 310 315 320

Cys Tyr Asn Thr His Glu Lys Arg Ile Tyr Gin Ser Asn Met Leu Asn

325 330 335

Arg Tyr Leu Ile Asn Val Thr Phe Glu Gly Arg Asn Leu Ser Phe Ser

340 345 350

Glu Asp Gly Tyr Gin Met His Pro Lys Leu Val Ile Ile Leu Leu Asn

355 360 365

Lys Glu Arg Lys Trp Glu Arg Val Gly Lys Trp Lys Asp Lys Ser Leu

370 375 380

Gin Met Lys Tyr Tyr Val Trp Pro Arg Met Cys Pro Glu Thr Glu Glu 385 390 395 400

Gin Glu Asp Asp His Leu Ser Ile Val Thr Leu Glu Glu Ala Pro Phe

405 410 415

Val Ile Val Glu Ser Val Asp Pro Leu Ser Gly Thr Cys Met Arg Asn

420 425 430

Thr Val Pro Cys Gin Lys Arg Ile Ile Ser Glu Asn Lys Thr Asp Glu

435 440 445

Glu Pro Gly Tyr Ile Lys Lys Cys Cys Lys Gly Phe Cys Ile Asp Ile

450 455 460

Leu Lys Lys Ile Ser Lys Ser Val Lys Phe Thr Tyr Asp Leu Tyr Leu 465 470 475 480

Val Thr Asn Gly Lys His Gly Lys Lys Ile Asn Gly Thr Trp Asn Gly

485 490 495

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

500 505 510

Leu Thr Ile Asn Glu Glu Arg Ser Glu Val Val Asp Phe Ser Val Pro

515 520 525

Phe Ile Glu Thr Gly Ile Ser Val Met Val Ser Arg Ser Asn Gly Thr

530 535 540

Val Ser Pro Ser Ala Phe Leu Glu Pro Phe Ser Ala Asp Val Trp Val 545 550 555 560

Met Met Phe Val Met Leu Leu Ile Val Ser Ala Val Ala Val Phe Val

565 570 575

Phe Glu Tyr Phe Ser Pro Val Gly Tyr Asn Arg Cys Leu Ala Asp Gly

580 585 590

Arg Glu Pro Gly Gly Pro Ser Phe Thr Ile Gly Lys Ala Ile Trp Leu 595 600 605 Leu Trp Gly Leu Val Phe Asn Asn Ser Val Pro Val Gin Asn Pro Lys

610 615 620

Gly Thr Thr Ser Lys Ile Met Val Ser Val Trp Ala Phe Phe Ala Val 625 630 635 640

Ile Phe Leu Ala Ser Tyr Thr Ala Asn Leu Ala Ala Phe Met Ile Gin

645 650 655

Glu Glu Tyr Val Asp Gin Val Ser Gly Leu Ser Asp Lys Lys Phe Gin

660 665 670

Arg Pro Asn Asp Phe Ser Pro Pro Phe Arg Phe Gly Thr Val Pro Asn

675 680 685

Gly Ser Thr Glu Arg Asn Ile Arg Asn Asn Tyr Ala Glu Met His Ala

690 695 700

Tyr Met Gly Lys Phe Asn Gin Arg Gly Val Asp Asp Ala Leu Leu Ser 705 710 715 720

Leu Lys Thr Gly Lys Leu Asp Ala Phe Ile Tyr Asp Ala Ala Val Leu

725 730 735

Asn Tyr Met Ala Gly Arg Asp Glu Gly Cys Lys Leu Val Thr Ile Gly

740 745 750

Ser Gly Lys Val Phe Ala Ser Thr Gly Tyr Gly Ile Ala Ile Gin Lys

755 760 765

Asp Ser Gly Trp Lys Arg Gin Val Asp Leu Ala Ile Leu Gin Leu Phe

770 775 780

Gly Asp Gly Glu Met Glu Glu Leu Glu Ala Leu Trp Leu Thr Gly Ile 785 790 795 800

Cys His Asn Glu Lys Asn Glu Val Met Ser Ser Gin Leu Asp Ile Asp

805 810 815

Asn Met Ala Gly Val Phe Tyr Met Leu Gly Ala Ala Met Ala Leu Ser

820 825 830

Leu Ile Thr Phe Ile Cys Glu His Leu Phe Tyr Trp Gin Phe Arg His

835 840 845

Cys Phe Met Gly Val Cys Ser Gly Lys Pro Gly Met Val Phe Ser Ile

850 855 860

Ser Arg Gly Ile Tyr Ser Cys Ile His Gly Val Ala Ile Glu Glu Arg 865 870 875 880

Gin Ser Val Met Asn Ser Pro Thr Ala Thr Met Asn Asn Thr His Ser

885 890 895

Asn Ile Leu Arg Leu Leu Arg Thr Ala Lys Asn Met Ala Asn Leu Ser

900 905 910

Gly Val Asn Gly Ser Pro Gin Ser Ala Leu Asp Phe Ile Arg Arg Glu

915 920 925

Ser Ser Val Tyr Asp Ile Ser Glu His Arg Arg Ser Phe Thr His Ser

930 935 940

Asp Cys Lys Ser Tyr Asn Asn Pro Pro Cys Glu Glu Asn Leu Phe Ser 945 950 955 960

Asp Tyr Ile Ser Glu Val Glu Arg Thr Phe Gly Asn Leu Gin Leu Lys 965 970 975 Asp Ser Asn Val Gin Asp His Tyr His His His His Arg Pro His

980 985 990

Ser Ile Gly Ser Thr Ser Ser Ile Asp Gly Leu Tyr Asp Cys Asp Asn

995 1000 1005

Pro Pro Phe Thr Thr Gin Pro Arg Ser Ile Ser Lys Lys Pro Leu Asp

1010 1015 1020

Ile Gly Leu Pro Ser Ser Lys His Ser Gin Leu Ser Asp Leu Tyr Gly 1025 1030 1035 1040

Lys Phe Ser Phe Lys Ser Asp Arg Tyr Ser Gly ^" His Asp Asp Leu Ile

1045 1050 1055

Arg Ser Asp Val Ser Asp Ile Ser Thr His Thr Val Thr Tyr Gly Asn

1060 1065 1070

Ile Glu Gly Asn Ala Ala Lys Arg Arg Lys Gin Gin Tyr Lys Asp Ser

1075 1080 1085

Leu Lys Lys Arg Pro Ala Ser Ala Lys Ser Arg Arg Glu Phe Asp Glu

1090 1095 1100

Ile Glu Leu Ala Tyr Arg Arg Arg Pro Pro Arg Ser Pro Asp His Lys 1105 1110 1115 1120

Arg Tyr Phe Arg Asp Lys Glu Gly Leu Arg Asp Phe Tyr Leu Asp Gin

1125 1130 1135

Phe Arg Thr Lys Glu Asn Ser Pro His Trp Glu His Val Asp Leu Thr

1140 1145 1150

Asp Ile Tyr Lys Glu Arg Ser Asp Asp Phe Lys Arg Asp Ser Val Ser

1155 1160 1165

Gly Gly Gly Pro Cys Thr Asn Arg Ser His Leu Lys His Gly Thr Gly

1170 1175 1180

Glu Lys His Gly Val Val Gly Gly Val Pro Ala Pro Trp Glu Lys Asn 1185 1190 1195 1200

Leu Thr Asn Val Asp Trp Glu Asp Arg Ser Gly Gly Asn Phe Cys Arg

1205 1210 1215

Ser Cys Pro Ser Lys Leu His Asn Tyr Ser Ser Thr Val Ala Gly Gin

1220 1225 1230

Asn Ser Gly Arg Gin Ala Cys Ile Arg Cys Glu Ala Cys Lys Lys Ala

1235 1240 1245

Gly Asn Leu Tyr Asp Ile Ser Glu Asp Asn Ser Leu Gin Glu Leu Asp

1250 1255 1260

Gin Pro Ala Ala Pro Val Ala Val Thr Ser Asn Ala Ser Ser Thr Lys 1265 1270 1275 1280

Tyr Pro Gin Ser Pro Thr Asn Ser Lys Ala Gin Lys Lys Asn Arg Asn

1285 1290 1295

Lys Leu Arg Arg Gin His Ser Tyr Asp Thr Phe Val Asp Leu Gin Lys

1300 1305 1310

Glu Glu Ala Ala Leu Ala Pro Arg Ser Val Ser Leu Lys Asp Lys Gly

1315 1320 1325

Arg Phe Met Asp Gly Ser Pro Tyr Ala His Met Phe Glu Met Pro Ala 1330 1335 1340 Gly Glu Ser Ser Phe Ala Asn Lys Ser Ser Val Pro Thr Ala Gly His 1345 1350 1355 1360

His His Asn Asn Pro Gly Ser Gly Tyr Met Leu Ser Lys Ser Leu Tyr

1365 1370 1375

Pro Asp Arg Val Thr Gin Asn Pro Phe Ile Pro Thr Phe Gly Asp Asp

1380 1385 1390

Gin Cys Leu Leu His Gly Ser Lys Ser Tyr phe Phe Arg Gin Pro Thr

1395 1400 1405

Val Ala Gly Ala Ser Lys Thr Arg Pro Asp Phe Arg Ala Leu Val Thr

1410 1415 1420

Asn Lys Pro Val Val Val Thr Leu His Gly Ala Val Pro Gly Arg Phe 1425 1430 1435 1440

Gin Lys Asp Ile Cys Ile Gly Asn Gin Ser Asn Pro Cys Val Pro Asn

1445 1450 1455

Asn Lys Asn Pro Arg Ala Phe Asn Gly Ser Ser Asn Gly His Val Tyr

1460 1465 1470

Glu Lys Leu Ser Ser Ile Glu Ser Asp Val

1475 1480

(2) INFORMATION ABOUT SEQ ID NO: 5: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 4738 base pairs

(B) TYPE: nucleic acid

(C) STRAND FORM: Single

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

(iii) HYPOTHETICAL: NO (iii) ANTISENSE: NO (vi) ORIGINAL ORIGIN:

(A) ORGANISM: Rattus norvegieus (F) FABRIC TYPE: Brain (ix) CHARACTERISTICS:

(A) NAME / KEY: CDS

(B) LOCATION: 674..3562

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5: GGGCGGCATC CAGGCCGGCC ACCCGCGCTG GTCCCGCCGC GCCCGGCTCC GGTGCCCGCC GAGCTCGCAG CCAGCGTGCC AGCTCGCGGG GGCTGGCGCT GGGACGAGCA CCGCCGGAGA 1 CTGTCGCGGG CTTTGGGCTG GGGTGAAGCG CCCTTCCCGA AGCCGCTGGG GCAAGGAATC 1 GGGGCAGGGG CAAGAGCCAG AGCCGAGGCT GGGTTTGACC AGAGCGCGGG GAGTTGGAAA 2 GGAGACAGAG CTCCGAGAGC AAAAAACCAG CTATAGCAGT TGGTGAGCTG GTCCCTGCTG 3 GGTACCTGCT GCTTCCACCG ACCCCCTCCC TCATTGGGGA TTTCCCCAGA CGCTGCCCCC 3 AACGTCTTGG TTCCCCTCAG CTCTCGTGCT CCAAAAGAAG GGTGGTAGGA AAAGGCCACG 4 GGAGAGCCTG CGCTCCGGTG CCCTGGGCCT GGAAGCACCG GTTGGCAGCT GTGGTCTCTT 4 GGGGCTTAAC CAGGACCCCC GACGGCTGAG AGGAGAAGCC CAAGCCTTTG GCTGCCGGAA 5 GGTTTTGTGC TTGGCCTAGG GAGGTTCTCT CTTCTCTGTC CTTGGCTGTG TGGTCTCTGC 6 CCTTCCTTCC TTCCTGTTGT CCATCTACCT CTCTCTATGC CTGCTCTGGT TCTCTGCAGA 6 CCCTCCAGGT GAC ATG GGT GGG GCC CTA GGG CCC GCC CTG CTG CTC ACT 709 Met Gly Gly Ala Leu Gly Pro Ala Leu Leu Leu Thr 1 5 10

TCA CTC CTT GGT GCT TGG GCA AGG CTG GGC GCA GGG CAG GGA GAA CAG 757 Ser Leu Leu Gly Ala Trp Ala Arg Leu Gly Ala Gly Gin Gly Glu Gin

15 20 25

GCC GTG ACT GTG GCG GTG GTG TTT GGC AGC TCT GGG CCA CTG CAG ACC 805 Ala Val Thr Val Ala Val Val Phe Gly Ser Ser Gly Pro Leu Gin Thr

30 35 -40

CAG GCC CGG ACT CGT CTT ACC TCC CAG AAC TTC CTG GAC CTG CCT CTG 853 Gin Ala Arg Thr Arg Leu Thr Ser Gin Asn Phe Leu Asp Leu Pro Leu 45 50 55 60

GAG ATC CAG CCA CTC ACC GTA GGG GTC AAC AAT ACC AAT CCC AGC AGC 901 Glu Ile Gin Pro Leu Thr Val Gly Val Asn Asn Thr Asn Pro Ser Ser

65 70 75

ATC CTC ACC CAA ATC TGC GGG CTT CTG GGT GCC GCC CGT GTC CAC GGC 949 Ile Leu Thr Gin Ile Cys Gly Leu Leu Gly Ala Ala Arg Val His Gly

80 85 90

ATC GTC TTT GAG GAC AAC GTG GAC ACC GAG GCC GTG GCC CAG CTA CTG 997 Ile Val Phe Glu Asp Asn Val Asp Thr Glu Ala Val Ala Gin Leu Leu

95 100 105

GAC TTC GTC TCC TCC CAG ACC CAC GTG CCC ATC CTC AGC ATC AGC GGA 1045 Asp Phe Val Ser Ser Gin Thr His Val Pro Ile Leu Ser Ile Ser Gly

110 115 120

GGC TCT GCT GTG GTC CTC ACC CCC AAG GAG CCC ACG TCC GCC TTC CTA 1093 Gly Ser Ala Val Val Leu Thr Pro Lys Glu Pro Thr Ser Ala Phe Leu 125 130 135 140

CAG CTG GGC GTG TCC CTG GAG CAG CAG CTG CAG GTG CTG TTC AAG GTG 1141 Gin Leu Gly Val Ser Leu Glu Gin Gin Leu Gin Val Leu Phe Lys Val

145 150 155

CTG GAG GAA TAC GAC TGG AGC GCG TTC GCG GTC ATC ACC AGC CTG CAT 1189 Leu Glu Glu Tyr Asp Trp Ser Ala Phe Ala Val Ile Thr Ser Leu His

160 165 170

CCG GGC CAC GCG CTC TTC CTC GAA GGC GTG CGC GCC GTC GCG GAC CGC 123 Pro Gly His Ala Leu Phe Leu Glu Gly Val Arg Ala Val Ala Asp Arg

175 180 185

AGC TAC CTG AGC TGG CGG CTG CTG GAC GTG CTC ACG CTG GAG CTA GGT 128 Ser Tyr Leu Ser Trp Arg Leu Leu Asp Val Leu Thr Leu Glu Leu Gly

190 195 200

CCG GGT GGG CCG CGA GCG CGC ACT CAG CCG CTG CTA CGC CAG GTC GAC 133 Pro Gly Gly Pro Arg Ala Arg Thr Gin Pro Leu Leu Arg Gin Val Asp 205 210 215 220

GCG CCG GTG CTG GTG GCT TAC TGC TCC CGT GAA GAG GCC GAG GTG CTC 138 Ala Pro Val Leu Val Ala Tyr Cys Ser Arg Glu Glu Ala Glu Val Leu 225 230 235 TTC GCT GAG GCT GCA CAG GCT GGC CTG GTG GGA CCC GGG CAC GTG TGG 142 Phe Ala Glu Ala Ala Gin Ala Gly Leu Val Gly Pro Gly His Val Trp

240 245 250

TTG GTA CCT AAC CTG GCG CTG GGA AGC ACC GAC GCA CCC CCT GCA GCT 147 Leu Val Pro Asn Leu Ala Leu Gly Ser Thr Asp Ala Pro Pro Ala Ala

255 260 265

TTC CCG GTG GGC CTC ATC AGT GTG GTC ACC GAG AGT TGG CGC CTT AGT 152 Phe Pro Val Gly Leu Ile Ser Val Val Thr Glu Ser Trp Arg Leu Ser

270 275 280

CTA CGC CAG AAG GTT CGC GAC GGA GTG GCC ATT CTG GCC CTC GGT GCC 157 Leu Arg Gin Lys Val Arg Asp Gly Val Ala Ile Leu Ala Leu Gly Ala 285 290 295 300

CAC AGC TAC CGA CGC CAG TAC GGT ACC CTG CCA GCC CCG GCT GGG GAC 162 His Ser Tyr Arg Arg Gin Tyr Gly Thr Leu Pro Ala Pro Ala Gly Asp

305 310 315

TGC CGA AGC CAC CCG GGA CCC GTC AGC CCT GCT AGG GAG GCC TTC TAC 166 Cys Arg Ser His Pro Gly Pro Val Ser Pro Ala Arg Glu Ala Phe Tyr

320 325 330

AGG CAT CTG CTG AAT GTC ACC TGG GAG GGC CGA GAC TTC TCT TTC AGC 171 Arg His Leu Leu Asn Val Thr Trp Glu Gly Arg Asp Phe Ser Phe Ser

335 340 345

CCT GGT GGG TAC CTG GTT CGG CCC ACC ATG GTT GTG ATC GCC CTC AAC 176 Pro Gly Gly Tyr Leu Val Arg Pro Thr Met Val Val Ile Ala Leu Asn

350 355 360

CGG CAC CGC CTC TGG GAG ATG GTG GGA CGG TGG GAC CAT GGA GTC CTG 181 Arg His Arg Leu Trp Glu Met Val Gly Arg Trp Asp His Gly Val Leu 365 370 375 380

TAC ATG AAG TAT CCA GTA TGG CCT CGC TAC AGC ACT TCT CTG CAG CCT 186 Tyr Met Lys Tyr Pro Val Trp Pro Arg Tyr Ser Thr Ser Leu Gin Pro

385 390 395

GTG GTG GAC AGC CGG CAC CTG ACA GTG GCC ACA CTG GAA GAA AGG CCC 190 Val Val Asp Ser Arg His Leu Thr Val Ala Thr Leu Glu Glu Arg Pro

400 405 410

TTT GTC ATT GTG GAG AGC CCT GAC CCT GGC ACA GGT GGC TGT GTT CCC 19 Phe Val Ile Val Glu Ser Pro Asp Pro Gly Thr Gly Gly Cys Val Pro

415 420 425

AAC ACC GTG CCC TGC CGC AGA CAG AGC AAC CAC ACC TTC AGC AGC GGG 20 Asn Thr Val Pro Cys Arg Arg Gin Ser Asn His Thr Phe Ser Ser Gly

430 435 440

GAT CTA ACC CCC TAC ACC AAG CTC TGT TGT AAG GGC TTC TGC ATC GAC 20 Asp Leu Thr Pro Tyr Thr Lys Leu Cys Cys Lys Gly Phe Cys Ile Asp 445 450 455 460

ATC CTC AAA AAG CTG GCC AAG GTG GTC AAG TTC TCC TAC GAC TTG TAC 21 Ile Leu Lys Lys Leu Ala Lys Val Val Lys Phe Ser Tyr Asp Leu Tyr 465 470 475 CTG GTG ACC AAC GGC AAA CAC GGC AAG AGG GTT CGT GGT GTG TGG AAC 2149 Leu Val Thr Asn Gly Lys His Gly Lys Arg Val Arg Gly Val Trp Asn

480 <: Ö 490

GGC ATG ATC GGG GAG GTA TAC TAC A-.G CGG GCA GAC ATG GCC ATC GGC 2197 Gly Met Ile Gly Glu Val Tyr Tyr Lys Arg Ala Asp Met Ala Ile Gly

495 500 505

TCC CTC ACC ATC AAT GAG GAG CGC TCT GAG ATT ATA GAT TTC TCT GTG 2245 Ser Leu Thr Ile Asn Glu Glu Arg Ser Glu Ile Ile Asp Phe Ser Val

510 515 520

CCT TTT GTG GAG ACC GGC ATC AGT GTT ATG GTA TCA AGG AGC AAC GGC 2293 Pro Phe Val Glu Thr Gly Ile Ser Val Met Val Ser Arg Ser Asn Gly 525 530 535 540

ACC GTC TCC CCC TCA GCT TTT CTG GAA CCC TAC AGC CCT GCC GTG TGG 234 Thr Val Ser Pro Ser Ala Phe Leu Glu Pro Tyr Ser Pro Ala Val Trp

545 550 555

GTG ATG ATG TTC GTG ATG TGT CTC ACG GTG GTT GCC ATC ACT GTC TTC 2389 Val Met Met Phe Val Met Cys Leu Thr Val Val Ala Ile Thr Val Phe

560 565 570

ATG TTC GAG TAC TTC AGC CCT GTC AGC TAC AAC CAG AAC CTC ACC AAG 243 Met Phe Glu Tyr Phe Ser Pro Val Ser Tyr Asn Gin Asn Leu Thr Lys

575 580 585

GGC AAG AAA CCT GGT GGA CCA TCT TTC ACC ATT GGC AAG TCC GTG TGG 248 Gly Lys Lys Pro Gly Gly Pro Ser Phe Thr Ile Gly Lys Ser Val Trp

590 595 600

TTG CTG TGG GCA CTG GTC TTC AAC AAC TCT GTT CCC ATC GAG AAC CCC 253 Leu Leu Trp Ala Leu Val Phe Asn Asn Ser Val Pro Ile Glu Asn Pro 605 610 615 620

CGG GGT ACC ACC AGC AAG ATC ATG GTC CTG GTG TGG GCC TTC TTC GCT 258 Arg Gly Thr Thr Ser Lys Ile Met Val Leu Val Trp Ala Phe Phe Ala

625 630 635

GTC ATC TTC CTC GCT AGC TAT ACG GCC AAT CTG GCG GCC TTC ATG ATC 262 Val Ile Phe Leu Ala Ser Tyr Thr Ala Asn Leu Ala Ala Phe Met Ile

640 645 650

CAG GAG CAA TAC ATC GAC ACT GTG TCG GGC CTT AGT GAC AAG AAG TTT 267 Gin Glu Gin Tyr Ile Asp Thr Val Ser Gly Leu Ser Asp Lys Lys Phe

655 660 665

CAG CGG CCT CAA GAC CAA TAC CCA CCC TTC CGT TTT GGC ACG GTA CCT 272 Gin Arg Pro Gin Asp Gin Tyr Pro Pro Phe Arg Phe Gly Thr Val Pro

670 675 680

AAC GGC AGC ACA GAG AGG AAC ATC CGT AGC AAC TAC GGT GAC ATG CAC 277 Asn Gly Ser Thr Glu Arg Asn Ile Arg Ser Asn Tyr Gly Asp Met His 685 690 695 700

ACC CAC ATG GTC AAG TTC AAC CAG CGC TCG GTG GAG GAC GCT CTG ACA 282 Thr His Met Val Lys Phe Asn Gin Arg Ser Val Glu Asp Ala Leu Thr 705 710 715 AGC CTG AAG ATG GGG AAG CTG GAC GCC TTC ATC TAT GAT GCT GCT GTG 286 Ser Leu Lys Met Gly Lys Leu Asp Ala Phe Ile Tyr Asp Ala Ala Val

720 725 730

CTC AAC TAC ATG GCG GGC AAG GAC GAA GGC TGC AAG CTG GTC ACC ATT 291 Leu Asn Tyr Met Ala Gly Lys Asp Glu Gly Cys Lys Leu Val Thr Ile

735 740 745

GGG TCT GGC AAA GTC TTT GCC ACC ACT GGC TAT GGC ATT GCC ATG CAG 296 Gly Ser Gly Lys Val Phe Ala Thr Thr Gly Tyr Gly Ile Ala Met Gin

750 755 760

AAA GAC TCC CAC TGG AAG CGG GCC ATA GAC CTG GCA CTC CTG CAA CTT 301 Lys Asp Ser His Trp Lys Arg Ala Ile Asp Leu Ala Leu Leu Gin Leu 765 770 775 780

CTG GGG GAT GGA GAG ACG CAG AAG CTG GAG ACA GTG TGG CTC TCA GGG 306 Leu Gly Asp Gly Glu Thr Gin Lys Leu Glu Thr Val Trp Leu Ser Gly

785 790 795

ATC TGC CAG AAC GAG AAG AAT GAG GTG ATG AGC AGC AAG CTG GAC ATT 310 Ile Cys Gin Asn Glu Lys Asn Glu Val Met Ser Ser Lys Leu Asp Ile

800 805 810

GAC AAC ATG GCG GGG GTC TTC TAC ATG CTG TTG GTG GCC ATG GGA CTG 315 Asp Asn Met Ala Gly Val Phe Tyr Met Leu Leu Val Ala Met Gly Leu

815 820 825

GCC CTT CTG GTC TTT GCC TGG GAG CAC CTG GTC TAC TGG AAA CTT CGA 320 Ala Leu Leu Val Phe Ala Trp Glu His Leu Val Tyr Trp Lys Leu Arg

830 835 840

CAC TCG GTG CCC AAC TCA TCC CAG CTG GAC TTC CTG CTG GCT TTC AGC 325 His Ser Val Pro Asn Ser Ser Gin Leu Asp Phe Leu Leu Ala Phe Ser 845 850 855 860

AGG GGC ATC TAC AGC TGC TTC AAC GGG GTA CAG AGC CTT CCG AGT CCC 330 Arg Gly Ile Tyr Ser Cys Phe Asn Gly Val Gin Ser Leu Pro Ser Pro

865 870 875

GCA CGG CCG CCC AGC CCG GAT CTC ACC GCA GAC TCA GCC CAG GCC AAT 334 Ala Arg Pro ^" Pro Ser Pro Asp Leu Thr Ala Asp Ser Ala Gin Ala Asn

880 885 890

GTG CTG AAG ATG CTG CAG GCG GCC CGA GAC ATG GTG AAC ACC GCG GAC 339 Val Leu Lys Met Leu Gin Ala Ala Arg Asp Met Val Asn Thr Ala Asp

895 900 905

GTG AGC AGC TCT TTG GAC CGC GCC ACT CGC ACC ATC GAG AAC TGG GGC 344 Val Ser Ser Ser Leu Asp Arg Ala Thr Arg Thr Ile Glu Asn Trp Gly

910 915 920

AAC AAC CGC CGC GTG CCT GCT CCC ACC GCC TCT GGC CCG CGG TCC TCC 34 Asn Asn Arg Arg Val Pro Ala Pro Thr Ala Ser Gly Pro Arg Ser Ser 925 930 935 940

ACC CCG GGT CCT CCG GGA CAA CCG AGC CCC AGC GGC TGG GGC CTC CTG 35 Thr Pro Gly Pro Pro Gly Gin Pro Ser Pro Ser Gly Trp Gly Leu Leu 945 950 955 GTG GGG GCC GCA CCC CGC TAGCGCGCAG GGCCCCGCAG CCTCCGGCTC 3589 Val Gly Ala Ala Pro Arg

960 GCCCCGCGAC GTGCGGGCCA CCTTTGCCCG ATGTGTCCCG ACCATCCTGC AGGCACGCTT 3649 CGGACGCGCG GTGGCCGGTG CGAGTTGGGC ATCAGGGACC GCACGTCTCG GCTTCCGAGC 3709 GGCGCGCGCT CCCGGAGCGC TCTCTGTTGC CCGCGCACTG CCACTACAGT TCCTTCCCTC 3769 GAGCGGAGAG ATCCGGGCGC CCCTACCTCC CGTTATTCCC GGAGCCCCCG GAGCCCGACG 3829 ACCTGCCTCT GCTCGGGCCG GAACAGCTGG CTCGGCGGGA AGCAATGCTG CGCGCGGCCT 3889 GGGCCCGGGG CCCGCGCGCT CGCCACGCTT CCTTGCCCAG CTCGGTGGTA GAAGCTTTCA 3949 CTCGATCCAA CCCTCTGCCT GCCAGGTGTA CCGGTCACGC CTGTGCCTGC CCGTGCCCCC 4009 AAAGCCGGCC ATCCTGCAGG CACTTGGCTC AGGCACAGTC GCTGAGGCTG CCGTCCTACC 4069 CGGCGGCCTG TGTGGAGGGC TGTACCAGCA TGGGGTGGCC ACCTGGCAGC CCAGACAGCA 4129 CGTCTGCCTG CACGCCCATA CCCGCCTGCC GTTCTGTTGG GGAACTGTCT GCCGTGACCC 4189 TCACCCTGTA CCAGCCACAG CCCCTGGCTC ATTGGAACCT GGGAGCCTCC AGCACACAGA 4249 GTCAGGACCC TGGGGCTAGG CACAGGCTAC AGGGACAGTG GGGTGCTGGA AGAGGTCAGC 4309 AGGGAAGCCT GTGGGACACA AGGTTTTCCA AGGTCCTGCA CCTGGAGGCG GGTCTCCAGC 4369 CTGGAATCAG AAGTGTGAGG TTACTGGCAG CTTTGGTTCC TGCTGGACTG GCTTCTGTGG 4429 CC GGCCTTGG CAGGTTTGGG GGACAGGCCA GCTTGGGCTC CTCTGGCTTC TATTTTGAAA 4489 TCCTGGCCAT GGACCCCAGT GAAAAGGATA TCTTCCAGGC CCAGCTTTTG ACTTAGTGTG 4549 GGCTGGGATC TTGTTCATCA CATGACCTCT TGCGGTGGTC ACAGGGAGTA AAAATGTGTC 4609 CCTGTCCTGT TGTCAGCTAG TTCCTAGTTG TGGTTGCTGT GGCCTTGGGT TGGGGACACA 4669 GAAATTGGAA ATCGAGGTTG GGGTGGGGCC ATCCCACTTC TTTGCGCCAA TAGCTGGGTT TCTTTTCAT 4729 4738 (2) INFORMATION FOR SEQ ID NO: 6 t

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 962 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) TYPE OF MOLECULE: Protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6: Met Gly Gly Ala Leu Gly Pro Ala Leu Leu Leu Thr Ser Leu Leu Gly

1 5 10 15

Ala Trp Ala Arg Leu Gly Ala Gly Gin Gly Glu Gin Ala Val Thr Val

20 25 30

Ala Val Val Phe Gly Ser Ser Gly Pro Leu Gin Thr Gin Ala Arg Thr

35 40 45

Arg Leu Thr Ser Gin Asn Phe Leu Asp Leu Pro Leu Glu Ile Gin Pro

50 55 60

Leu Thr Val Gly Val Asn Asn Thr Asn Pro Ser Ser Ile Leu Thr Gin 65 70 75 80

Ile Cys Gly Leu Leu Gly Ala Ala Arg Val His Gly Ile Val Phe Glu

85 90 95

Asp Asn Val Asp Thr Glu Ala Val Ala Gin Leu Leu Asp Phe Val Ser

100 105 110

Ser Gin Thr His Val Pro Ile Leu Ser Ile Ser Gly Gly Ser Ala Val 115 120 125 Val Leu Thr Pro Lys Glu Pro Thr Ser Ala Phe Leu Gin Leu Gly Val

130. 135 140

Ser Leu Glu Gin Gin Leu Gin Val Leu Phe Lys Val Leu Glu Glu Tyr 145 150 155 160

Asp Trp Ser Ala Phe Ala Val Ile Thr Ser Leu His Pro Gly His Ala

165 170 175

Leu Phe Leu Glu Gly Val Arg Ala Val Ala Asp Arg Ser Tyr Leu Ser

180 185 190

Trp Arg Leu Leu Asp Val Leu Thr Leu Glu Leu Gly Pro Gly Gly Pro

195 200 205

Arg Ala Arg Thr Gin Pro Leu Leu Arg Gin Val Asp Ala Pro Val Leu

210 215 220

Val Ala Tyr Cys Ser Arg Glu Glu Ala Glu Val Leu Phe Ala Glu Ala 225 230 235 240

Ala Gin Ala Gly Leu Val Gly Pro Gly His Val Trp Leu Val Pro Asn

245 250 255

Leu Ala Leu Gly Ser Thr Asp Ala Pro Pro Ala Ala Phe Pro Val Gly

260 265 270

Leu Ile Ser Val Val Thr Glu Ser Trp Arg Leu Ser Leu Arg Gin Lys

275 280 285

Val Arg Asp Gly Val Ala Ile Leu Ala Leu Gly Ala His Ser Tyr Arg

290 295 300

Arg Gin Tyr Gly Thr Leu Pro Ala Pro Ala Gly Asp Cys Arg Ser His 305 310 315 320

Pro Gly Pro Val Ser Pro Ala Arg Glu Ala Phe Tyr Arg His Leu Leu

325 330 335

Asn Val Thr Trp Glu Gly Arg Asp Phe Ser Phe Ser Pro Gly Gly Tyr

340 345 350

Leu Val Arg Pro Thr Met Val Val Ile Ala Leu Asn Arg His Arg Leu

355 360 365

Trp Glu Met Val Gly Arg Trp Asp His Gly Val Leu Tyr Met Lys Tyr

370 375 380

Pro Val Trp Pro Arg Tyr Ser Thr Ser Leu Gin Pro Val Val Asp Ser 385 390 395 400

Arg His Leu Thr Val Ala Thr Leu Glu Glu Arg Pro Phe Val Ile Val

405 410 415

Glu Ser Pro Asp Pro Gly Thr Gly Gly Cys Val Pro Asn Thr Val Pro

420 425 430

Cys Arg Arg Gin Ser Asn His Thr Phe Ser Ser Gly Asp Leu Thr Pro

435 440 445

Tyr Thr Lys Leu Cys Cys Lys Gly Phe Cys Ile Asp Ile Leu Lys Lys

450 455 460

Leu Ala Lys Val Val Lys Phe Ser Tyr Asp Leu Tyr Leu Val Thr Asn 465 470 475 480

Gly Lys His Gly Lys Arg Val Arg Gly Val Trp Asn Gly Met Ile Gly 485 490 495 Glu Val Tyr Tyr Lys Arg Ala Asp Met Ala Ile Gly Ser Leu Thr Ile

500. 505 510

Asn Glu Glu Arg Ser Glu Ile Ile Asp Phe Ser Val Pro Phe Val Glu

515 520 525

Thr Gly Ile Ser Val Met Val Ser Arg Ser Asn Gly Thr Val Ser Pro

530 535 540

Ser Ala Phe Leu Glu Pro Tyr Ser Pro Ala Val Trp Val Met Met Phe 545 550 555 560

Val Met Cys Leu Thr Val Val Ala Ile Thr Val Phe Met Phe Glu Tyr

565 570 575

Phe Ser Pro Val Ser Tyr Asn Gin Asn Leu Thr Lys Gly Lys Lys Pro

580 585 590

Gly Gly Pro Ser Phe Thr Ile Gly Lys Ser Val Trp Leu Leu Trp Ala

595 600 605

Leu Val Phe Asn Asn Ser Val Pro Ile Glu Asn Pro Arg Gly Thr Thr

610 615 620

Ser Lys Ile Met Val Leu Val Trp Ala Phe Phe Ala Val Ile Phe Leu 625 630 635 640

Ala Ser Tyr Thr Ala Asn Leu Ala Ala Phe Met Ile Gin Glu Gin Tyr

645 650 655

Ile Asp Thr Val Ser Gly Leu Ser Asp Lys Lys Phe Gin Arg Pro Gin

660 665 670

Asp Gin Tyr Pro Pro Phe Arg Phe Gly Thr Val Pro Asn Gly Ser Thr

675 680 685

Glu Arg Asn Ile Arg Ser Asn Tyr Gly Asp Met His Thr His Met Val

690 695 700

Lys Phe Asn Gin Arg Ser Val Glu Asp Ala Leu Thr Ser Leu Lys Met 705 710 715 720

Gly Lys Leu Asp Ala Phe Ile Tyr Asp Ala Ala Val Leu Asn Tyr Met

725 730 735

Ala Gly Lys Asp Glu Gly Cys Lys Leu Val Thr Ile Gly Ser Gly Lys

740 745 750

Val Phe Ala Thr Thr Gly Tyr Gly Ile Ala Met Gin Lys Asp Ser His

755 760 765

Trp Lys Arg Ala Ile Asp Leu Ala Leu Leu Gin Leu Leu Gly Asp Gly

770 775 780

Glu Thr Gin Lys Leu Glu Thr Val Trp Leu Ser Gly Ile Cys Gin Asn 785 790 795 800

Glu Lys Asn Glu Val Met Ser Ser Lys Leu Asp Ile Asp Asn Met Ala

805 810 815

Gly Val Phe Tyr Met Leu Leu Val Ala Met Gly Leu Ala Leu Leu Val

820 825 830

Phe Ala Trp Glu His Leu Val Tyr Trp Lys Leu Arg His Ser Val Pro

835 840 845

Asn Ser Ser Gin Leu Asp Phe Leu Leu Ala Phe Ser Arg Gly Ile Tyr 850 855 860 Ser Cys Phe Asn Gly Val Gin Ser Leu Pro Ser Pro Ala Arg Pro Pro 865 870 875 880

Ser Pro Asp Leu Thr Ala Asp Ser Ala Gin Ala Asn Val Leu Lys Met

885 890 895

Leu Gin Ala Ala Arg Asp Met Val Asn Thr Ala Asp Val Ser Ser Ser

900 905 910

Leu Asp Arg Ala Thr Arg Thr Ile Glu Asn Trp Gly Asn Asn Arg Arg

915 920 925

Val Pro Ala Pro Thr Ala Ser Gly Pro Arg Ser Ser Thr Pro Gly Pro

930 935 940

Pro Gly Gin Pro Ser Pro Ser Gly Trp Gly Leu Leu Val Gly Ala Ala 945 950 955 960

Per arg

(2) INFORMATION ABOUT SEQ ID NO: 7: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 18 base pairs

(B) TYPE: nucleic acid

(C) STRAND FORM: Single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iii) ANTISENSE: NO (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7: AATTCCATGG ATGCATGC 1

(2) INFORMATION ABOUT SEQ ID NO: 8: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 27 base pairs

(B) TYPE: nucleic acid

(C) STRAND FORM: Single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iii) ANTISENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8: GCGAATTCTG GAAYGGMTGA TGGGNGA 2

(2) INFORMATION ABOUT SEQ ID NO: 9: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRAND FORM: Single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iii) ANTISENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 9: GCGGTACCAA RGCDCCARRT TDGCWGTRT (2) INFORMATION ON SEQ ID NO: 10: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 8 amino acids

(B) TYPE: amino acid

(C) STRAND FORM: Single

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

(iii) HYPOTHETICAL: NO (iii) ANTISENSE: NO

(v) FRAGMENT TYPE: internal (xi) SEQÜENZBESCHREIBÜNG ^': SEQ ID NO: 10: Trp Asn Gly Gly Glu His His Ile 1 5

(2) INFORMATION ON SEQ ID NO: 11: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 8 amino acids

(B) TYPE: amino acid

(C) STRAND FORM: Single

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

(iii) HYPOTHETICAL: NO (iii) ANTISENSE: NO

(v) TYPE OF FRAGMENT: inner (i) SEQUENCE DESCRIPTION: SEQ ID NO: 11: Tyr Thr Ala Asn Leu Ala Ala Phe 1 5

(2) INFORMATION ON SEQ ID NO: 12: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 11 base pairs

(B) TYPE: nucleic acid

(C) STRAND FORM: Single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iii) ANTISENSEr NO (i) SEQUENCE DESCRIPTION: SEQ ID NO: 12: CTTAGAGCAC A 1

Claims

1. DNA sequences that code for NMDA receptor subunits and that from the group "that of

a) DNA sequences of the structure described in SEQ ID NO 1, 3 or 5,

b) DNA sequences which code for proteins with the structure described in SEQ ID NO 2, 4 or 6, and

c) DNA sequences which hybridize under standard conditions with DNA sequences a) or b) and do not code for NR1 are formed,

are selected.

2. Process for the genetic engineering production of NMDA receptor subunits using DNA sequences according to claim 1.

3. Use of DNA sequences according to claim 1 for identifying functional ligands for NMDA receptors.

4. A method for identifying functional ligands for NMDA receptors, characterized in that cells are transfected with a DNA sequence coding for an NMDA receptor according to claim 1, the membranes of these cells are isolated and conventional receptor binding experiments are carried out with these membranes.

5. A method for identifying functional ligands for NMDA receptors, characterized in that cells are transfected with a DNA sequence coding for an NMDA receptor according to claim 1 and the change in the cells caused by binding of the ligands to the receptor in these cells se¬ cond messenger mirror detected by a reporter system. Subunits of NMDA receptors, processes for their production and their use

Summary

The invention relates to new subunits for NMDA receptors and DNA sequences coding for them, and to production processes for DNA sequences and receptors. The invention further relates to methods for identifying functional ligands for this receptor.