NZ527682A - Protein Cluster V - Google Patents

Abstract

An isolated nucleic acid molecule comprises the sequence shown in SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17 or 19 and encodes polypeptides referred to as Protein Cluster V and useful in diagnosis of metabolic diseases. Also described are vectors, host cells and methods for producing the polypeptides.

Description

New Zealand Paient Spedficaiion for Paient Number 527682 PROTEIN CLUSTER V 5276 82 TECHNICAL FIELD The present invention relates to the identification of a human gene family expressed in metabolically relevant tissues. The genes encode a group polypeptides referred to as "Protein Cluster V" which are predicted to be useful in the diagnosis of metabolic diseases, such as obesity and diabetes, as well as in the identification of agents useful in the treatment of the said diseases.

BACKGROUND ART Metabolic diseases are defined as any of the diseases or disorders that disrupt normal metabolism. They may arise from nutritional deficiencies; in connection with diseases of the endocrine system, the liver, or the kidneys; or as a result of genetic defects. Metabolic diseases are conditions caused by an abnormality in one or more of the chemical reactions essential to producing energy, to regenerating cellular constituents, or to eliminating unneeded products arising from these processes. Depending on which metabolic pathway is involved, a single defective chemical reaction may producc consequences that are narrow, involving a single body function, or broad, affecting many organs and systems.

One of the major hormones that influence metabolism is insulin, which is synthesized in the beta cells of the islets of Langerhans of the pancreas. Insulin primarily regulates the direction of metabolism, shifting many processes toward the storage of substrates and away from their degradation. Insulin acts to increase the transport of glucose and amino acids as well as key minerals such as potassium, magnesium, and phosphate from the blood into cells. It also regulates a variety of enzymatic reactions within the cells, all of which have a common overall direction, namely the synthesis of large molecules from small units. A deficiency in the action of insulin (diabetes mellitus) causes severe impairment in (i) the storage of glucose in the form of glycogen and the oxidation of glucose for energy; (ii) the synthesis and storage of fat from fatty acids and their precursors and the completion of fatty-acid oxidation; and (iii) the synthesis of proteins from amino acids.

There are two varieties of diabetes. Type I is insulin-dependent diabetes mellitus (IDDM), for which insulin injection is required; it was formerly referred to as juvenile onset diabetes. In this type, insulin is not secreted by the pancreas and hence must be taken by injection. Type II, non-insulin-dependent diabetes mellitus (NIDDM) may be controlled by dietary restriction. It derives from insufficient pancreatic insulin secretion and tissue resistance to secreted insulin, which is complicated by subtle changes in the secretion of insulin by the beta cells. Despite their former classifications as juvenile or adult, either type can occur at any age; NIDDM, however, is the most common type, accounting for 90 percent of all diabetes. While the exact causes of diabetes remain obscure, it is evident that NIDDM is linked to heredity and obesity. There is clearly a genetic predisposition to NIDDM diabetes in those who become overweight or obese.

Obesity is usually defined in terms of the body mass index (BMI), i.e. weight (in kilograms) divided by the square of the height (in meters). Weight is regulated with great precision. Regulation of body weight is believed to occur not only in persons of normal weight but also among many obese persons, in whom obesity is attributed to an elevation in the set point around which weight is regulated. The determinants of obesity can be divided into genetic, environmental, and regulatory.

Recent discoveries have helped explain how genes may determine obesity and how they may influence the regulation of body weight. For example, mutations in the oh gene have led to massive obesity in mice. Cloning the ob gene led to the identification of leptin, a protein coded by this gene; leptin is produced in adipose tissue cells and acts to control body fat. The existence of leptin supports the idea that body weight is regulated, because leptin serves as a signal between adipose tissue and the areas of the brain that control energy metabolism, which influences body weight.

Metabolic diseases like diabetes and obesity are clinically and genetically heterogeneous disorders. Recent advances in molecular genetics have led to the recognition of genes involved in IDDM and in some subtypes of NIDDM, including maturity-onset diabetes of the young (MODY) (Velho & Froguel (1997) Diabetes Metab. 23 Suppl 2:34-37). However, several IDDM susceptibility genes have not yet been identified, and very little is known about genes contributing to common forms of NIDDM. Studies of candidate genes and of genes mapped in animal models of IDDM or NIDDM, as well as whole genome scanning of diabetic families from different populations, should allow the identification of most diabetes susceptibility genes and of the molecular targets for new potential drugs. The identification of genes involved in metabolic disorders will thus contribute to the development of novel predictive and therapeutic approaches.

The p3-adrenergic receptor (AR) represents one of a number of potential anti-obesity drugs targets for which selective agonists have been developed. In rodents, p3-AR mRNA is abundant in white adipose tissue (WAT) and brown adipose tissue (BAT). It has been demonstrated that mice lacking endogenous p3-adrenoceptors have a slight increase in body fat, but otherwise appear normal (Susulic V.S., et al. (1995) J. Biol. Chem. 270(49): 29483-29492). These mice are completely resistant to the specific p3-agonist CL-316,243, which has been shown to increase lipolysis, energy expenditure and affect insulin and leptin levels. When the (33-AR was ectopically expressed in white and brown adipose tissue or brown adipose tissue only, it was recently demonstrated that the anorectic and insulin secretagogue effects appeared to be mediated by white adipose tissue (Grujic D, et al. (1997) J Biol Chem. 272(28): 17686-93). How these effects are mediated by P3-AR agonists remains poorly understood.

Lardizabal, K.D. et al. (J. Biol. Chem. 276: 38862-38869) and Cases, S. et al. (J. Biol. Chem. 276: 38870-38876; both papers published 31 July 2001) disclose a new gene family, including members in fungi, plants and animals, which encode proteins corresponding to the "Cluster V" proteins according to the present invention. The proteins were shown to have acyl CoA:diacylglycerol acyltransferase (DGAT; EC 2.3.1.20) function. The gene family is unrelated to the previously identified DGAT(l) family and was designated DGAT2. DGAT2 was shown to have high expression levels in liver and white adipose tissue, suggesting that it may play a significant role in mammalian triglyceride metabolism.

DISCLOSURE OF THE INVENTION According to the present invention, a family of genes and encoded homologous proteins (hereinafter referred to as "Protein Cluster V") has been identified. Consequently, the present invention provides an isolated nucleic acid molecule selected from: (a) nucleic acid molecules comprising a nucleotide sequence as shown in SEQ ID NO: 3,5,7,9,11,13, 15,17, or 19. (b) nucleic acid molecules comprising a nucleotide sequence capable of hybridizing, under stringent hybridization conditions, to a nucleotide sequence complementary to the polypeptide coding region of a nucleic acid molecule as defined in (a); and (c) nucleic acid molecules comprising a nucleic acid sequence which is degenerate as a result of the genetic code to a nucleotide sequence as defined in (a) or (b).

The nucleic acid molecules according to the present invention includes cDNA, chemically synthesized DNA, DNA isolated by PCR, genomic DNA, and combinations thereof. RNA transcribed from DNA is also encompassed by the present invention.

The term "stringent hybridization conditions" is known in the art from standard protocols (e.g. Ausubel et al., supra) and could be understood as e.g. hybridization to filter-bound DNA in 0.5 M NaHPO^ 7% sodium dodecyl sulfate (SDS), 1 mM EDTA at +65°C, and washing in O.lxSSC / 0.1% SDS at +68°C.

I n a preferred form of the invention, the said nucleic acid molecule has a nucleotide sequence identical with SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17, or 19 of the Sequence Listing. However, the nucleic acid molecule according to the invention is not to be limited strictly to the sequence shown as SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17, or 19. Rather the invention encompasses nucleic acid molecules carrying modifications like substitutions, small deletions, insertions or inversions, which nevertheless encode proteins having substantially the features of the Protein Cluster V polypeptide according to the invention. Included in the invention are consequently nucleic acid molecules, the nucleotide sequence of which is at least 90% homologous, preferably at least 95% homologous, 'with the nucleotide sequence shown as SEQ ID NO: 3, 5, 7, 9,11, 13, 15, 17, or 19 in the Sequence Listing.

Included in the invention is also a nucleic acid molecule which nucleotide sequence is degenerate, because of the genetic code, to the nucleotide sequence shown as SEQ ID NO: 3, 5,7,9, 11, 13,15,17, or 19. A sequential grouping of three nucleotides, a "codon", codes for one amino acid. Since there are 64 possible codons, but only 20 natural amino acids, most amino acids are coded for by more than one codon. This natural "degeneracy", or "redundancy", of the genetic code is well known in the art. It will thus be appreciated that the nucleotide sequence shown in the Sequence Listing is only an example within a large but definite group of sequences which will encode the Protein Cluster V polypeptide.

The nucleic acid molecules according to the invention have numerous applications in techniques known to those skilled in the art of molecular biology. These techniques include their use as hybridization probes, for chromosome and gene mapping, in PCR technologies, in the production of sense or antisense nucleic acids, in screening for new therapeutic molecules, etc.

More specifically, the sequence information provided by the invention makes possible large-scale expression of the encoded polypeptides by techniques well known in the art. Nucleic acid molecules of the invention also permit identification and isolation of nucleic acid molecules encoding related polypeptides, such as human allelic variants and species homologues, by well-known techniques including Southern and/or Northern hybridization, and PCR. Knowledge of the sequence of a human DNA also makes possible, through use of Southern hybridization or PCR, the identification of genomic DNA sequences encoding the proteins in Cluster V, expression control regulatory sequences such as promoters, operators, enhancers, repressors, and the like. Nucleic acid molecules of the invention are also useful in hybridization assays to detect the capacity of cells to express the proteins in Cluster V. Nucleic acid molecules of the invention may also provide a basis for diagnostic methods useful for identifying a genetic aiteration(s) in a locus that underlies a disease state or states, which information is useful both for diagnosis and for selection of therapeutic strategies.

In a further aspect, the invention provides an isolated polypeptide encoded by the nucleic acid molecule as defined above. In a preferred form, the said polypeptide has an amino acid sequence according to SEQ ID NO: 4, 6, 8, 10, 12, 14,16, 18 or 20 of the Sequence Listing. However, the polypeptide according to the invention is not to be limited strictly to a polypeptide with an amino acid sequence identical with SEQ ID NO: 4, 6, 8, 10,12,14,16, 18 or 20 in the Sequence Listing. Rather the invention encompasses polypeptides carrying modifications like substitutions, small deletions, insertions or inversions, which polypeptides nevertheless have substantially the features of the Protein Cluster V polypeptide. Included in the invention are consequently polypeptides, the amino acid sequence of which is at least 90% homologous, preferably at least 95% homologous, with the amino acid sequence shown as SEQ ID NO: 4, 6, 8, 10,12, 14,16, 18 or 20 in the Sequence Listing.

In a further aspect, the invention provides a vector harboring the nucleic acid molecule as defined above. The said vector can e.g. be a replicable expression vector, which carries and is capable of mediating the expression of a DNA molecule according to the invention. In the present context the term "replicable" means that the vector is able to replicate in a given type of host cell into which is has been introduced. Examples of vectors are viruses such as bacteriophages, cosmids, plasmids and other recombination vectors. Nucleic acid molecules are inserted into vector genomes by methods well known in the art.

Included in the invention is also a cultured host cell harboring a vector according to the invention. Such a host cell can be a prokaryotic cell, a unicellular eukaryotic cell or a cell derived from a multicellular organism. The host cell can thus e.g. be a bacterial cell such as an E. coli cell; a cell from yeast such as Saccharomyces cervisiae or Pichia - 7 — pastoris, or a mammalian cell. The methods employed to effect introduction of the vector into the host cell are standard methods well known to a person familiar with recombinant DNA methods.

In yet another aspect, the invention provides a process for production of a polypeptide, comprising culturing a host cell, according to the invention, under conditions whereby said polypeptide is produced, and recovering said polypeptide. The medium used to grow the cells may be any conventional medium suitable for the purpose. A suitable vector may be any of the vectors described above, and an appropriate host cell may be any of the cell types listed above. The methods employed to construct the vector and effect introduction thereof into the host cell may be any methods known for such purposes within the field of recombinant DNA. The recombinant polypeptide expressed by the cells may be secreted, i.e. exported through the cell membrane, dependent on the type of cell and the composition of the vector.

In a further aspect, the invention provides a method for identifying an agent capable of modulating a nucleic acid molecule according to the invention, comprising (i) providing a cell comprising the said nucleic acid molecule; (ii) contacting said cell with a candidate agent; and (iii) monitoring said cell for an effect that is not present in the absence of said candidate agent.

For screening purposes, appropriate host cells can be transformed with a vector having a reporter gene under the control of the nucleic acid molecule according to this invention. The expression of the reporter gene can be measured in the presence or absence of an agent with known activity (i.e. a standard agent) or putative activity (i.e. a "test agent" or "candidate agent"). A change in the level of expression of the reporter gene in the presence of the test agent is compared with that effected by the standard agent. In this way, active agents are identified and their relative potency in this assay determined.

A transfection assay can be a particularly useful screening assay for identifying an effective agent. In a transfection assay, a nucleic acid containing a gene such as a - 8 — reporter gene that is operably linked to a nucleic acid molecule according to the invention, is transfected into the desired cell type. A test level of reporter gene expression is assayed in the presence of a candidate agent and compared to a control level of expression. An effective agent is identified as an agent that results in a test level of expression that is different than a control level of reporter gene expression, which is the level of expression determined in the absence of the agent. Methods for transfecting cells and a variety of convenient reporter genes are well known in the art (see, for example, Goeddel (ed.), Methods Enzymol., Vol. 185, San Diego: Academic Press, Inc. (1990); see also Sambrook, supra).

Throughout this description the terms "standard protocols" and "standard procedures", when used in the context of molecular biology techniques, are to be understood as protocols and procedures found in an ordinary laboratory manual such as: Current Protocols in Molecular Biology, editors F. Ausubel et al., John Wiley and Sons, Inc. 1994, or Sambrook, J., Fritsch, E.F. and Maniatis, T., Molecular Cloning: A laboratory manual, 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY 1989.

EXAMPLES EXAMPLE 1: Identification of protein clusters A family of homologous proteins (hereinafter referred to as "Protein Cluster V") was identified by an "all-versus-all" BLAST procedure using all Caenorhabditis elegans proteins in the Wormpep20 database release (http://www.sanger.ac.uk/Projects/ C_elegans/wormpep/index.shtml). The Wormpep database contains the predicted proteins from the C. elegans genome sequencing project, carried out jointly by the Sanger Centre in Cambridge, UK and the Genome Sequencing Center in St. Louis, USA. A number of 18,940 proteins were retrieved from Wormpep20. The proteins were used in a Smith-Waterman clustering procedure to group together proteins of similarity (Smith T.F. & Waterman M.S. (1981) Identification of common molecular subsequences. J. Mol. Biol. 147(1): 195-197; Pearson WR. (1991) Searching protein sequence libraries: comparison of the sensitivity and selectivity of the Smith-Waterman and FASTA algorithms. Genomics 11: 635-650; Olsen et al. (1999) Optimizing Smith-Waterman alignments. Pac Symp Biocomput.302-313). Completely annotated proteins were filtered out, whereby 10,130 proteins of unknown function could be grouped into 1,800 clusters.

The obtained sequence clusters were compared to the Drosophila melanogaster proteins contained in the database Flybase (Berkeley Drosophila Genome Project; http://www.fruitfly.org), and annotated clusters were removed. Non-annotated protein clusters, conserved in both C. elegans and D. melanogaster, were saved to a worm/fly data set, which was used in a BLAST procedure (http://www.nchi.nlm.nih.gov/ Education/BLASTinfo/information3.hlml) against the Celera Human Genome Database (http://www.celera.com). Overlapping fragments were assembled to, as close as possible, full-length proteins using the PHRAP software, developed at the University of Washington (http://www.genome.washington.edu/ UWGC/analysistools/phrap.htm). A group of homologous proteins ("Protein Cluster V") with unknown function was chosen for further studies.

EST databases provided by the EMBL (http://www.embl.org/Services/index.html) were used to check whether the human proteins in Cluster V were expressed, in order to identify putative pseudogenes. One putative pseudogene was identified and excluded.

EXAMPLE 2: Analyses of Protein Cluster V (a) Alignment The human part of this protein family includes seven different 150-250 residue polypeptides shown as SEQ ID NO: 2, 4, 6, 8, 10, 12, 14,16,18 and 20, encoded by the nucleic acid sequences shown as SEQ ID NO: 1, 3, 5, 7, 9,11, 13, 15, 17, and 19. The amino acid sequence shown as SEQ ID NO: 2 was identified to correspond to a human 261 aa sequence encoded by the gene "WUGSC: H_DJ0747G18.5" (GenBank Accession No. AC004876). No function has been associated with the said gene.

An alignment of the human polypeptides included in Protein Cluster V, using the ClustalW multiple alignment software (Thompson et al. (1994) Nucleic Acid Research 22: 4673-4680) is shown in Table I. The alignment showed a high degree of conservation over a 100 residues region in the protein (corresponding to positions 23-147 in SEQ ID NO: 2), indicating the presence of a novel domain. (b) HMM-Pfam A HMM-Pfam search was performed on the human family members. Pfam is a large collection of protein families and domains. Pfam contains multiple protein alignments and profile-HMMs (Profile Hidden Markov Models) of these families. Profile-HMMs can be used to do sensitive database searching using statistical descriptions of a sequence family's consensus. Pfam is available on the WWW at http://pfcim.wiisll.edu; http://www.sanger.ac.uk/Software/PJdm; and http://www.cgr.ki.se/Pfam. The latest version (4.3) of Pfam contains 1815 families. These Pfam families match 63% of proteins in SWISS-PROT 37 and TrEMBL 9. For references to Pfam, see Bateman et al. (2000) The Pfam protein families database. Nucleic Acids Res. 28:263-266; Sonnhammer et al. (1998) Pfam: Multiple Sequence Alignments and HMM-Profiles of Protein Domains. Nucleic Acids Research, 26:322-325; Sonnhammer et al. (1997) Pfam: a Comprehensive Database of Protein Domain Families Based on Seed Alignments. Proteins 28: 405-420.

The HMM-Pfam search indicated that no previously known domains could be identified in Protein Cluster V. (c) TM-HMM The human proteins in Cluster V were analyzed using the TM-HMM tool available e.g. at http://wwM>. cbs.dtu.dk/services/TMHMM-1.0. TM-HMM is a method to model and predict the location and orientation of alpha helices in membrane-spanning proteins (Sonnhammer et al. (1998) A hidden Markov model for predicting transmembrane helices in protein sequences. ISMB 6:175-182). The results indicate that the human Cluster V proteins contain 3-4 transmembrane segments. (d) Analysis of non-human orthologs The Caenorhabditis elegans genome includes four genes, designated K07B1.4 (GenBank Accession No. AF003384), F59A1.10 (GenBank Accession No. Z81557), Y53G8B.2 (GenBank Accession No. AC006804), and W01A11.2 (GenBank Accession No. U64852) orthologous to the human Cluster V genes. The closest ancestor (K07B1.4) is on average 44% identical to the 10 human gene products. (See also: Genome sequence of the nematode C. elegans: a platform for investigating biology; The C. elegans Sequencing Consortium. Science (1998) 282:2012-2018. Published errata appear in Science (1999) 283:35; 283:2103; and 285:1493.) The Drosophila melanogaster genome includes four genes orthologous to human Cluster V. The most closely related genes, designated "CG1942" (GenBank Accession No. AE003840_36) and gene: "CGI 946" (GenBank Accession No. AE003840_37) are 39% identical to the human gene products. (See also Adams et al. (2000) The genome sequence of Drosophila melanogaster, Science 287:2185-2195) is 42% identical to the human protein set.

The human proteins in Cluster V show 27% identity to two yeast proteins; S. Cerevisiae SCYOR245CJ (GenBank Accession No. Z75153) and S. pombe SPCC548J (GenBank Accession No. AL359685). The yeast proteins are of unknown function.

EXAMPLE 3: Expression analysis The tissue distribution of the human genes was studied using the Incyte LifeSeq® database (http://www.incyte.com). The genes shown as SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15,17 were found to be expressed primarily in the following tissues: SEQ ID NO: 1 and 3: Liver, digestive system SEQ ID NO: 7 and 9: Exocrine Glands, Connective Tissue, Germ Cells SEQ ID NO: 11: Female genitalia, urinary tract SEQ ID NO: 17: Female genitalia, nervous system SEQ ID NO: 13 and 15: Digestive System SEQ ID NO: 5: Cardiovascular system Therefore, the said nucleic acid molecules and the encoded polypeptides shown are proposed to be useful for differential identification of the tissues or cell types present in a biological sample and for diagnosis of diseases and disorders related to the tissues where the genes are expressed.

EXAMPLE 4: Effect of (33-AR agonists on cluster V genes.

Microarrays consist of a highly ordered matrix of thousands of different DNA sequences that can be used to measure DNA and RNA variation in applications that include gene expression profiling, comparative genomics and genotyping (For recent reviews, see e.g.: Harrington et al. (2000) Monitoring gene expression using DNA microarrays. Curr. Opin. Microbiol. 3(3): 285-291; or Duggan etal. (1999) Expression profiling using cDNA Microarrays. Nature Genetics Supplement 21:10-14).

In order to investigate the mechanisms whereby P3-AR agonists affect gene regulation in adipose tissue in vivo, a study was carried out using Affymetrix GeneChip oligonucleotide arrays by comparing the transcript profiles of a large number of genes in white adipose tissue derived from C57BL/6J mice treated with the p3-AR agonist CL-316,243, or from control mice injected with a saline solution.

PolyA+mRNAs were extracted from white adipose tissue from control and 03-AR agonist treated mice respectively. They were reverse transcribed using a T7-tagged oligo-dT primer and double-stranded cDNAs were generated. These cDNAs were then amplified and labeled using In Vitro Transcription (fVT) with T7 RNA polymerase and biotinylated nucleotides. The populations of cRNAs obtained after IVT were purified and fragmented by heat to produce a distribution of RNA fragment sizes from approximately 35 to 200 bases. Two Affymetrix Mul 9K and Mul IK sets of 3 arrays (subA, subB and subC) and 2 arrays (subA and subB) respectively, were hybridized (using the recommended buffer) overnight at 45°C with the control or the treated denatured samples. The arrays were then washed and stained with R-phycoerythrin streptavidin with the help of an Affymetrix fluidics station. The cartridges were scanned using a Hewlett-Packard confocal scanner and the images were analyzed with the GeneChip 3.1 software (Affymetrix).

The results indicate that the mouse gene (GenBank accession No. AA275948), orthologous to the worm gene F59A1.10, is down-regulated by P3-AR agonist treatment. It is hypothesized that the human genes in Cluster V are similarly involved in metabolically important signaling pathways.

EXAMPLE 5: Multiple Tissue Northern blotting Multiple Tissue Northern blotting (MTN) is performed to make a more thorough analysis of the expression profiles of the proteins in Cluster V. Multiple Tissue Northern (MTN™) Blots (http://www.clontech.com/mtn) are pre-made Northern blots featuring Premium Poly A+ RNA from a variety of different human, mouse, or rat tissues. MTN Blots can be used to analyze size and relative abundance of transcripts in different tissues. MTN Blots can also be used to investigate gene families and alternate splice forms and to assess cross species homology.

EXAMPLE 6: Identification of polypeptides binding to Protein Cluster V In order to assay for proteins interacting with Protein Cluster V, the two-hybrid screening method can be used. The two-hybrid method, first described by Fields & Song (1989) Nature 340:245-247, is a yeast-based genetic assay to detect protein-protein interactions in vivo. The method enables not only identification of interacting proteins, but also results in the immediate availability of the cloned genes for these proteins.

The two-hybrid method can be used to determine if two known proteins (i.e. proteins for which the corresponding genes have been previously cloned) interact. Another important application of the two-hybrid method is to identify previously unknown proteins that interact with a target protein by screening a two-hybrid library. For reviews, see e.g.: Chien et al. (1991) The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. Proc. Natl. Acad. Sci. U.S.A. 88:9578-9582; Bartel PL, Fields (1995) Analyzing protein-protein interactions using two-hybrid system. Methods Enzymol. 254:241-263; or Wallach et al. (1998) The yeast two-hybrid screening technique and its use in the study of protein-protein interactions in apoptosis. Curr. Opin. Immunol. 10(2): 131-136. See also http://www. clontech. com/matchmaker.

The two-hybrid method uses the restoration of transcriptional activation to indicate the interaction between two proteins. Central to this technique is the fact that many eukaryotic transcriptional activators consist of two physically discrete modular domains: the DNA-binding domain (DNA-BD) that binds to a specific promoter sequence and the activation domain (AD) that directs the RNA polymerase II complex to transcribe the gene downstream of the DNA binding site. The DNA-BD vector is used to generate a fusion of the DNA-BD and a bait protein X, and the AD vector is used to generate a fusion of the AD and another protein Y. An entire library of hybrids with the AD can also be constructed to search for new or unknown proteins that interact with the bait protein. When interaction occurs between the bait protein X and a candidate protein Y, the two functional domains, responsible for DNA binding and activation, are tethered, resulting in functional restoration of transcriptional activation. The two hybrids are cotransformed into a yeast host strain harboring reporter genes containing appropriate upstream binding sites; expression of the reporter genes then indicates interaction between a candidate protein and the target protein.

EXAMPLE 7: Full-length cloning of Cluster V genes The polymerase chain reaction (PCR), which is a well-known procedure for in vitro enzymatic amplification of a specific DNA segment, can be used for direct cloning of Protein Cluster V genes. Tissue cDNA can be amplified by PCR and cloned into an appropriate plasmid and sequenced. For reviews, see e.g. Hooft van Huijsduijnen (1998) PCR-assisted cDNA cloning: a guided lour of the minefield. Biotechniques 24:390-392; Lenstra (1995) The applications of the polymerase chain reaction in the life sciences. Cellular & Molecular Biology 41:603-614; or Rashtchian (1995) Novel methods for cloning and engineering genes using the polymerase chain reaction. Current Opinion in Biotechnology 6:30-36. Various methods for generating suitable ends to facilitate the direct cloning of PCR products are given e.g. in Ausubel et al. supra (section 15.7).

In an alternative approach to isolate a cDNA clone encoding a full length protein of Protein Cluster V, a DNA fragment corresponding to a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17 or 19, or aportion thereof, can be used as a probe for hybridization screening of a phage cDNA library. The DNA fragment is amplified by the polymerase chain reaction (PCR) method. The primers are preferably 10 to 25 nucleotides in length and are determined by procedures well known to those skilled in the art. A lambda phage library containing cDNAs cloned into lambda phage-vectors is plated on agar plates with E. coli host cells, and grown. Phage plaques are transferred to nylon membranes, which are hybridized with a DNA probe prepared as described above. Positive colonies are isolated from the plates. Plasmids containing cDNA are rescued from the isolated phages by standard methods. Plasmid DNA is isolated from the clones. The size of the insert is determined by digesting the plasmid with appropriate restriction enzymes. The sequence of the entire insert is determined by automated sequencing of the plasmids.

EXAMPLE 8: Recombinant expression of proteins in eukaryotic host cells To produce proteins of Cluster V, a polypeptide-encoding nucleic acid molecule is expressed in a suitable host cell using a suitable expression vector and standard genetic engineering techniques. For example, the polypeptide-encoding sequence is subcloned into a commercial expression vector and transfected into mammalian, e.g. Chinese Hamster Ovary (CHO), cells using a standard transfection reagent. Cells stably expressing a protein are selected. Optionally, the protein may be purified from the cells using standard chromatographic techniques. To facilitate purification, antisera is raised against one or more synthetic peptide sequences that correspond to portions of the amino acid sequence, and the antisera is used to affinity purify the protein.

EXAMPLE 9: Determination of gene function Methods are known in the art for elucidating the biological function or mode of action of individual genes. For instance, RNA interference (RNAi) offers a way of specifically and potently inactivating a cloned gene, and is proving a powerful tool for investigating gene function. For reviews, see e.g. Fire (1999) RNA-triggeredgene silencing. Trends in Genetics 15:358-363; or Kuwabara & Coulson (2000) RNAi-praspects for a general technique for determining gene function. Parasitology Today 16:347-349. When double-stranded RNA (dsRNA) corresponding to a sense and antisense sequence of an endogenous mRNA is introduced into a cell, the cognate mRNA is degraded and the gene is silenced. This type of posttranscriptional gene silencing (PTGS) was first discovered in C. elegans (Fire et al., (1998) Nature 391:806-811). RNA interference has recently been used for targeting nearly 90% of predicted genes on C. elegans chromosome I (Fraser et al. (2000) Nature 408: 325-330) and 96% of predicted genes on C. elegans chromosome III (Gonczy et al. (2000) Nature 408:331-336).

TABLE I Alignment of polypeptides in Protein Cluster V SEQ_2 SEQ_4 SEQ_8 SEQ_10 SEQ_i2 SEQ_14 SEQ_20 MVNGKSITSLQSNKNLAAIHGPKYLCGNFGPRWQAFSLGTKLDPMEVFPKLLPSKVPVAQ 60 SEQ_16 SEQ_18 SEQ 6 SEQ_2 SEQ_4 SEQ_8 SEQ_10 SEQ_12 SEQ 14 SEQ_20 TLAPYSAPCFQRLWWSAAKVKAPSHNAKQGPKMDGQLVKTHDLSPKHNYIIANHPHGILS 120 SEQ_16 RPGGSEG 7 SEQ_18 SEQ_6 SEQ_2 EAPLFSRCLAFHPPFILLNTPKLVKTAELPPDRNYVLGAHPHGIMCTGFLCNF 53 SEQ_4 LGTLLGWRAPLFSRCLAFHPPFILLNTPKLVKTAELPPDRNYVLGAHPHGIMCTGFLCNF 60 SEQ_8 —AFCNFSTEATEVSKKFPGIRPYLATLAGNFRMPVLREYLMSGGICPVSRDTIDYLLSK 58 SEQ_!0 SEQ_12 SEQ_14 NLF 3 SEQ_20 FGVFINFATEATGIARIFPSITPFVGTLERIFWIPIVREYVMSMGVCPVSSSALKYLLTQ 180 SEQ_16 RFPKVTPVSGRVRAGTQAPPWLSRLPSLQLVKTAELDPSRNYIAGFHPHGVLAVGAFANL 67 SEQ_18 SDYVPLKLLKTHDICPSRNYILVCHPHGLFAHGWFGHF 38 SEQ_6 CSEIFASLRLPR 1MAHSKQPSHFQSLMLLQW 31 SEQ_2 STESHGFSQLFPGLRPWLSVLAG LFYLPVYRDYIMSFGLCPVSRQSLD FIL 104 SEQ_4 STESNGFSQLFPGLRPWLAVLAG LFYLPVYRDYIMSFGASLVPVYSFGENDIFRL 115 SEQ_8 NGSGNAIIIVVGGAAESLSSMPGKNAVTLRNRKGFVKLALRHGADLVPIYSFGENEVYKQ 118 SEQ_10 RNRKG FVKLALRHGADLVPI YSFGENEVYKQ 31 SEQ_12 KESLDAHPGKFTLFIRQRKGFVKIALTHGASLVPVVSFGENELFKQ 4 6 S EQ_14 EAHKLKFN11VGGAQEALDARPGS FTLLLRNRKGFVRLALT HGAPLVXIFS FG EN DL FDQ 63 SEQ_20 KGSGNAVVIVVGGAAEALLCRPGASTLFLKQRKGFVKMALQTGAYLVPSYSFGENEVFNQ 24 0 SEQ_16 CTESTGFSSIFPGIRPHLMMLTL WFRAPFFRDYIMSAGLVTSEKESAAHILNRKG 122 SEQ_18 ATEASGFSKIFPGITPYILTLGA FFWMPFLREYVMSTGACSVSRSSIDFLLTHKG 93 GEQ 6 PLSYLAIFWILQPLFVYLLFTSLWPLPVLYFAWLFLDWKTPERGGRRSAWVRNWCVWTHI 91

Claims (15)

WO 02/083721 -18- PCT/SE02/00730 TABLE 1 (continued) SEQ_2 SQPQLG QAVVI MVGGAEALYSVPGEHCLTLQKRKGFVRLALRHGASLVP 153 SEQ_4 KAFATGSWQHWCQLTFKK LMGFSPCIFWGRGLFSATSWGLLPFAVPITTVVGRPIP 171 SEQ_8 VIFEEGSWGRWVQKKFQ KYIGFAPCIFHGRGLFSSDTWGLVPYSKPITTVVGEPIT 17 4 SEQ_10 VIFEEGSWGRWVQKKFQ KYIGFAPCIFHGRGLFSSDTWGLVPYSKPITTVGGGKIQ 87 SEQ_12 T DNPEGSWIRTVQNKLQ KIMGFALPLFHARGVFQYN-FGLMTYRKAIHTVVGRPIP 101 SEQ_14 IPNSSGSWLRYIQNRLQ KIMG 84 SEQ_20 ETFPEGTWLRLFQKTFQDTFKKILGLNFCTFHGRG-FTRGSWGFLPFNRPITTVVGEPLP 299 SEQ_16 GGNLLGIIVG GAQEALDARPGSFTLLLRNRKGFVRLALTHG 163 SEQ_18 TGNMVIVVIG GLAECRYSLPGSSTLVLKNRSGFVRMALQHGVPLIP 139 SEQ 6 RDYFPITILK TKDLSPEHNYLMGVHPMGLLTFGAFCNFC 130 SEQ_2 VYS FGENDIFRLtfAFATGSWQHWCQLTFKKL-MGFSPCIFWVAV 196 SEQ_4 VPQRLHPTEEEVNHYHALYMTDLEQLFEEHKESCGVPASTCLTFI-- 216 SEQ_8 IPKLEHPTQQDIDLYHTMYMEALVKLFDKHKTKFGLPETEVLEVN-- 219 SEQ_10 S RSKKRKINXX QNDSCYSL 106 SEQ_12 VRQTLNPTQEQIEELHQTYMEELRKLFEEHKGKYGIPEHETLVLK— 14 6 SEQ_14 SEQ_20 IPRIKRPNQKTVDKYHALYISALRKLFDQHKVEYGLPETQELTIT-- 344 SEQ_16 SEQ_18 AYAFGETDL 148 SEQ 6 WO 02/083721 CLAIMS - 19- PCT/SE02/00730

1. An isolated nucleic acid molecule selected from: (a) nucleic acid molecules comprising a nucleotide sequence as shown in SEQ ID NO: 3, 5, 7, 9,11,13,15,17, or 19; (b) nucleic acid molecules comprising a nucleotide sequence capable of hybridizing, under stringent hybridization conditions, to a nucleotide sequence complementary to the polypeptide coding region of a nucleic acid molecule as defined in (a); and (c) nucleic acid molecules comprising a nucleic acid sequence which is degenerate as a result of the genetic code to a nucleotide sequence as defined in (a) or (b).

2. An isolated polypeptide encoded by the nucleic acid molecule according to claim 1.

3. The isolated polypeptide according to claim 2 having an amino acid sequence shown as SEQ ID NO: 4, 6, 8, 10,12, 14, 16, 18 or 20 in the Sequence Listing

4. A vector harboring the nucleic acid molecule according to claim 1.

5. A replicable expression vector which carries and is capable of mediating the expression of a nucleotide sequence according to claim 1.

6. A cultured host cell harboring a vector according to claim 4 or 5.

7. A process for production of a polypeptide, comprising culturing a host cell according to claim 6 under conditions whereby said polypeptide is produced, and recovering said polypeptide.

8. A method for identifying an agent capable of modulating a nucleic acid molecule according to claim 1, comprising (i) providing a cell comprising the said nucleic acid molecule; -20- (ii) contacting said cell with a candidate agent; and (iii) monitoring said cell for an effect that is not present in the absence of said candidate agent.

9. An isolated nucleic acid as claimed in claim 1 substantially as herein described with reference to any example thereof.

10. An isolated polypeptide as claimed in claim 2 substantially as herein described with reference to any example thereof.

11. A vector as claimed in claim 4 substantially as herein described with reference to any example thereof.

12. A replicable expression vector as claimed in claim 5 substantially as herein described with reference to any example thereof.

13. A cultured host cell as claimed in claim 6 substantially as herein described with reference to any example thereof.

14. A process as claimed in claim 7 substantially as herein described with reference to any example thereof.

15. A method as claimed in claim 8 substantially as herein described with reference to any example thereof. 253033_1.DOC WO 02/083721 PCT/SE02/00730 - 1 - SEQUENCE LISTING <110> Pharmacia AB <120> Protein Cluster V <130> 00407 <160> 20 <170> Patentln version 3.0 <210> 1 <211> 593 <212> DNA <213> human <220> <221> CDS <222> (3) . . (593) <400> 1 tg gag gcc cct ctt ttc age egg tgt ctt gee ttc cat cct ccc ttc 47 Glu Ala Pro Leu Phe Ser Arg Cys Leu Ala Phe His Pro Pro Phe 15 10 15 ate ctg ctc aac acc ccg aag ctg gtg aaa aca gca gag ctg ccc ccg 95 lie Leu Leu Asn Thr Pro Lys Leu Val Lys Thr Ala Glu Leu Pro Pro 20 25 30 gat egg aac tac gtg ctg ggc gcc cac cct cat ggg ate atg tgt aca 143 Asp Arg Asn Tyr Val Leu Gly Ala His Pro His Gly lie Met Cys Thr 35 40 45 ggc ttc ctc tgt aat ttc tcc acc gag age cat ggc ttc tcc cag ctc 191 Gly Phe Leu Cys Asn Phe Ser Thr Glu Ser His Gly Phe Ser Gin Leu 50 55 60 ttc ccg ggg ctc egg ccc tgg tta tcc gtg ctg get ggc ctc ttc tac 239 Phe Pro Gly Leu Arg Pro Trp Leu Ser Val Leu Ala Gly Leu Phe Tyr 65 70 75 ctc ccg gtc tat cgc gac tac ate atg tcc ttt gga ctc tgt ccg gtg 287 Leu Pro Val Tyr Arg Asp Tyr He Met Ser Phe Gly Leu Cys Pro Val 80 85 90 95 age cgc cag age ctg gac ttc ate ctg tcc cag ccc cag ctc ggg cag 335 Ser Arg Gin Ser Leu Asp Phe lie Leu Ser Gin Pro Gin Leu Gly Gin 100 105 110 gcc gtg gtc ate atg gtg ggg ggt gcg cac gag gcc ctg tat tea gtc 383 Ala Val Val lie Met Val Gly Gly Ala His Glu Ala Leu Tyr Ser Val 115 120 125 ccc ggg gag cac tgc ctt acg ctc cag aag cgc aaa ggc ttc gtg cgc 431 Pro Gly Glu His Cys Leu Thr Leu Gin Lys Arg Lys Gly Phe Val Arg 130 135 140 WO 02/083721 -2- PCT/SE02/00730 ctg gcg ctg agg cac ggg gcg tcc ctg gtg ccc gtg tac tcc ttt ggg 479 Leu Ala Leu Arg His Gly Ala Ser Leu Val Pro Val Tyr Ser Phe Gly 145 150 155 gag aat gac ate ttt aga ctt aag get ttt gcc aca ggc tcc tgg cag 527 Glu Asn Asp lie Phe Arg Leu Lys Ala Phe Ala Thr Gly Ser Trp Gin 160 165 170 175 cat tgg tgc cag ctc acc ttc aag aag ctc atg ggc ttc tct cct tgc 575 His Trp Cys Gin Leu Thr Phe Lys Lys Leu Met Gly Phe Ser Pro Cys 180 185 190 ate ttc tgg gtc gcg gtc 593 He Phe Trp Val Ala Val 195 <210> 2 <211> 197 <212> PRT <213> human <4 00> 2 Glu Ala Pro Leu Phe Ser Arg Cys Leu Ala Phe His Pro Pro Phe lie 1 5 10 15 Leu Leu Asn Thr Pro Lys Leu Val Lys Thr Ala Glu Leu Pro Pro Asp 20 25 30 Arg Asn Tyr Val Leu Gly Ala His Pro His Gly lie Met Cys Thr Gly 35 40 45 Phe Leu Cys Asn Phe Ser Thr Glu Ser His Gly Phe Ser Gin Leu Phe 50 55 60 Pro Gly Leu Arg Pro Trp Leu Ser Val Leu Ala Gly Leu Phe Tyr Leu 65 70 75 80 Pro Val Tyr Arg Asp Tyr lie Met Ser Phe Gly Leu Cys Pro Val Ser 85 90 95 Arg Gin Ser Leu Asp Phe lie Leu Ser Gin Pro Gin Leu Gly Gin Ala 100 105 110 Val Val lie Met Val Gly Gly Ala His Glu Ala Leu Tyr Ser Val Pro 115 120 125 Gly Glu His Cys Leu Thr Leu Gin Lys Arg Lys Gly Phe Val Arg Leu 130 135 140 Ala Leu Arg His Gly Ala Ser Leu Val Pro Val Tyr Ser Phe Gly Glu 145 150 155 160 Asn Asp lie Phe Arg Leu Lys Ala Phe Ala Thr Gly Ser Trp Gin His 165 170 175 Trp Cys Gin Leu Thr Phe Lys Lys Leu Met Gly Phe Ser Pro Cys lie 180 185 190 WO 02/083721 -3- PCT/SE02/00730 Phe Trp Val Ala Val 195 <210> 3 <211> 822 <212> DNA <213> human <220> <221> CDS <222> (93)..(740) <400> 3 aaaaaaaaac ctgggccctt aaccctatcc taagaacctt taactcggaa ctctgctggg 60 gtggcccttg accctatcct aagaaccttt aa ctc gga act ctg ttg ggg tgg 113 Leu Gly Thr Leu Leu Gly Trp 1 5 agg gcc cct ctt ttc age egg tgt ctt gcc ttc cat cct ccc ttc ate 161 Arg Ala Pro Leu Phe Ser Arg Cys Leu Ala Phe His Pro Pro Phe lie 10 15 20 ctg ctc aac acc ccg aag ctg gtg aaa aca gca gag ctg ccc ccg gat 209 Leu Leu Asn Thr 'Pro Lys Leu Val Lys Thr Ala Glu Leu Pro Pro Asp 25 30 35 egg aac tac gtg ctg ggc gcc cac cct cat ggg ate atg tgt aca ggc 257 Arg Asn Tyr Val Leu Gly Ala His Pro His Gly lie Met Cys Thr Gly 40 45 50 55 ttc ctc tgt aat ttc tcc acc gag age aat ggc ttc tcc cag ctc ttc 305 Phe Leu Cys Asn Phe Ser Thr Glu Ser Asn Gly Phe Ser Gin Leu Phe 60 65 70 ccg ggg ctc egg ccc tgg tta gcc gtg ctg get ggc ctc ttc tac ctc 353 Pro Gly Leu Arg Pro Trp Leu Ala Val Leu Ala Gly Leu Phe Tyr Leu 75 80 85 ccg gtc tat cgc gac tac ate atg tcc ttt ggg gcg tcc ctg gtg ccc 401 Pro Val Tyr Arg Asp Tyr lie Met Ser Phe Gly Ala Ser Leu Val Pro 90 95 100 gtg tac tcc ttt ggg gag aat gac ate ttt aga ctt aag get ttt gcc 449 Val Tyr Ser Phe Gly Glu Asn Asp lie Phe Arg Leu Lys Ala Phe Ala 105 110 115 aca ggc tcc tgg cag cat tgg tgc cag ctc acc ttc aag aag ctc atg 4 97 Thr Gly Ser Trp Gin His Trp Cys Gin Leu Thr Phe Lys Lys Leu Met 120 125 130 135 ggc ttc tct cct tgc ate ttc tgg ggt cgc ggt ctc ttc tea gcc acc 545 Gly Phe Ser Pro Cys lie Phe Trp Gly Arg Gly Leu Phe Ser Ala Thr 140 145 150 tcc tgg ggc ctg ctg ccc ttt get gtg ccc ate acc act gtg gtg ggc 593 Ser Trp Gly Leu Leu Pro Phe Ala Val Pro lie Thr Thr Val Val Gly 155 160 165 WO 02/083721 -4 - PCT/SE02/00730 cgc ccc ate ccc gtc ccc cag cgc ctc cac ccc acc gag gag gaa gtc 641 Arg Pro lie Pro Val Pro Gin Arg Leu His Pro Thr Glu Glu Glu Val 170 175 180 aat cac tat cac gcc ctc tac atg acg gac ctg gag cag ctc ttc gag 689 Asn His Tyr His Ala Leu Tyr Met Thr Asp Leu Glu Gin Leu Phe Glu 185 190 195 gag cac aag gaa age tgt ggg gtc ccc get tcc acc tgc ctc acc ttc 737 Glu His Lys Glu Ser Cys Gly Val Pro Ala Ser Thr Cys Leu Thr Phe 200 205 210 215 ate taggcctggc cgcggccttt cgctgagccc ctgagcccaa ggcactgaga 7 90 lie cctccaccca ctgtggactc catgcctcca at 822 <210> 4 <211> 216 <212> PRT <213> human <4 00> 4 Leu Gly Thr Leu Leu Gly Trp Arg Ala Pro Leu Phe Ser Arg Cys Leu 15 10 15 Ala Phe His Pro Pro Phe lie Leu Leu Asn Thr Pro Lys Leu Val Lys 20 25 30 Thr Ala Glu Leu Pro Pro Asp Arg Asn Tyr Val Leu Gly Ala His Pro 35 40 45 His Gly lie Met Cys Thr Gly Phe Leu Cys Asn Phe Ser Thr Glu Ser 50 55 60 Asn Gly Phe Ser Gin Leu Phe Pro Gly Leu Arg Pro Trp Leu Ala Val 65 70 75 80 Leu Ala Gly Leu Phe Tyr Leu Pro Val Tyr Arg Asp Tyr lie Met Ser 85 90 95 Phe Gly Ala Ser Leu Val Pro Val Tyr Ser Phe Gly Glu Asn Asp lie 100 105 110 Phe Arg Leu Lys Ala Phe Ala Thr Gly Ser Trp Gin His Trp Cys Gin 115 120 125 Leu Thr Phe Lys Lys Leu Met Gly Phe Ser Pro Cys lie Phe Trp Gly 130 135 140 Arg Gly Leu Phe Ser Ala Thr Ser Trp Gly Leu Leu Pro Phe Ala Val 145 150 155 160 Pro lie Thr Thr Val Val Gly Arg Pro lie Pro Val Pro Gin Arg Leu 165 170 175 His Pro Thr Glu Glu Glu Val Asn His Tyr His Ala Leu Tyr Met Thr 180 185 190 WO 02/083721 -5- PCT/SE02/00730 Asp Leu Glu Gin Leu Phe Glu Glu His Lys Glu Ser Cys Gly Val Pro 195 200 205 Ala Ser Thr Cys Leu Thr Phe lie 210 215 <210> 5 <211> 392 <212> DNA <213> human <220> <221> CDS <222> (3)..(392) <4 00> 5 ac tgt tct gag ate ttt gcc tcc ctc agg ctc ccg aga ate atg get 47 Cys Ser Glu lie Phe Ala Ser Leu Arg Leu Pro Arg lie Met Ala 15 10 15 cat tec aag cag cct agt cac ttc cag agt ctg atg ctt ctg cag tgg 95 His Ser Lys Gin Pro Ser His Phe Gin Ser Leu Met T.eu Leu Gin Trp 20 25 30 cct ttg age tac ctt gcc ate ttt tgg ate ttg cag cca ttg ttc gtc 143 Pro Leu Ser Tyr Leu Ala lie Phe Trp lie Leu Gin Pro Leu Phe Val 35 40 45 tac ctg ctg ttt aca tcc ttg tgg ccg eta cca gtg ctt tac ttt gcc 191 Tyr Leu Leu Phe Thr Ser Leu Trp Pro Leu Pro Val Leu Tyr Phe Ala 50 55 60 tgg ttg ttc ctg gac tgg aag acc cca gag cga ggt ggc agg cgt teg 239 Trp Leu Phe Leu Asp Trp Lys Thr Pro Glu Arg Gly Gly Arg Arg Ser 65 70 75 gcc tgg gta agg aac tgg tgt gtc tgg acc cac ate agg gac tat ttc 287 Ala Trp Val Arg Asn Trp Cys Val Trp Thr His lie Arg Asp Tyr Phe 80 85 90 95 ccc att acg ate ctg aag aca aag gac eta tea cct gag cac aac tac 335 Pro lie Thr lie Leu Lys Thr Lys Asp Leu Ser Pro Glu His Asn Tyr 100 105 110 ctc atg ggg gtt cac ccc atg ggc ctc ctg acc ttt ggc gcc ttc tgc 383 Leu Met Gly Val His Pro Met Gly Leu Leu Thr Phe Gly Ala Phe Cys 115 120 125 aac ttc tgc 392 Asn Phe Cys 130 <210> 6 <211> 130 <212> PRT <213> human WO 02/083721 -6- PCT/SE02/00730 <400> 6 Cys Ser Glu lie Phe Ala Ser Leu Arg Leu Pro Arg lie Met Ala His 15 10 15 Ser Lys Gin Pro Ser His Phe Gin Ser Leu Met Leu Leu Gin Trp Pro 20 25 30 Leu Ser Tyr Leu Ala lie Phe Trp lie Leu Gin Pro Leu Phe Val Tyr 35 40 45 Leu Leu Phe Thr Ser Leu Trp Pro Leu Pro Val Leu Tyr Phe Ala Trp 50 55 60 Leu Phe Leu Asp Trp Lys Thr Pro Glu Arg Gly Gly Arg Arg Ser Ala 65 70 75 80 Trp Val Arg Asn Trp Cys Val Trp Thr His lie Arg Asp Tyr Phe Pro 85 90 95 lie Thr lie Leu Lys Thr Lys Asp Leu Ser Pro Glu His Asn Tyr Leu 100 105 110 Met Gly Val His Pro Met Gly Leu Leu Thr Phe Gly Ala Phe Cys Asn 115 120 125 Phe Cys 130 <210> 7 <211> 2519 <212> DNA <213> human <220> <221> CDS <222> (714)..(1373) <4 00> 7 gccgcctctg ctggggtcta ggctgtttct ctcgcgccac cactggccgc cggccgcagc 60 tccaggtgtc ctagccgccc agcctcgacg ccgtcccggg acccctgtgc tctgcgcgaa 120 gccctggccc cgggggccgg ggcatgggcc aggggcgcgg ggtgaagcgg cttcccgcgg 180 ggccgtgact gggcgggctt cagccatgaa gaccctcata gccgcctact ccggggtcct 240 gcgcggcgag cgtcaggccg aggctgaccg gagccagcgc tctcacggag gacctgcgct 300 gtcgcgcgag gggtctggga gatggggcac tggatccagc atcctctccg ccctccagga 360 cctcttctct gtcacctggc tcaataggtc caaggtggaa aagcagctac aggtcatctc 420 agtgctccag tgggtcctgt ccttccttgt actgggagtg gcctgcagtg ccatcctcat 480 gtacatattc tgcactgatt gctggctcat cgctgtgctc tacttcactt ggctggtgtt 540 tgactggaac acacccaaga aaggtggcag gaggtcacag tgggtccgaa actgggctgt 600 gtggcgctac tttcgagact actttcccat ccagctggtg aagacacaca acctgctgac 660 WO 02/083721 -7- PCT/SE02/00730 caccaggaac tatatctttg gataccaccc ccatggtatc atgggcctgg get gcc 716 Ala 1 ttc tgc aac ttc age aca gag gcc aca gaa gtg age aag aag ttc cca 7 64 Phe Cys Asn Phe Ser Thr Glu Ala Thr Glu Val Ser Lys Lys Phe Pro 5 10 15 ggc ata egg cct tac ctg get aca ctg gca ggc aac ttc cga atg cct 812 Gly lie Arg Pro Tyr Leu Ala Thr Leu Ala Gly Asn Phe Arg Met Pro 20 25 30 gtg ttg agg gag tac ctg atg tct gga ggt ate tgc cct gtc age egg 860 Val Leu Arg Glu Tyr Leu Met Ser Gly Gly lie Cys Pro Val Ser Arg 35 40 45 gac acc ata gac tat ttg ctt tea aag aat ggg agt ggc aat get ate 908 Asp Thr lie Asp Tyr Leu Leu Ser Lys Asn Gly Ser Gly Asn Ala lie 50 55 60 65 ate ate gtg gtc ggg ggt gcg get gag tct ctg age tcc atg cct ggc 956 lie lie Val Val Gly Gly Ala Ala Glu Ser Leu Ser Ser Met Pro Gly 70 75 80 aag aat gca gtc acc ctg egg aac cgc aag ggc ttt gtg aaa ctg gcc 1004 Lys Asn Ala Val Thr Leu Arg Asn Arg Lys Gly Phe Val Lys Leu Ala 85 90 95 ctg cgt cat gga get gac ctg gtt ccc ate tac tcc ttt gga gag aat 1052 Leu Arg His Gly Ala Asp Leu Val Pro lie Tyr Ser Phe Gly Glu Asn 100 105 110 gaa gtg tac aag cag gtg ate ttc gag gag ggc tcc tgg ggc cga tgg 1100 Glu Val Tyr Lys Gin Val lie Phe Glu Glu Gly Ser Trp Gly Arg Trp 115 120 125 gtc cag aag aag ttc cag aaa tac att ggt ttc gcc cca tgc ate ttc 1148 Val Gin Lys Lys Phe Gin Lys Tyr lie Gly Phe Ala Pro Cys lie Phe 130 135 140 145 cat ggt cga ggc ctc ttc tcc tcc gac acc tgg ggg ctg gtg ccc tac 1196 His Gly Arg Gly Leu Phe Ser Ser Asp Thr Trp Gly Leu Val Pro Tyr 150 155 160 tcc aag ccc ate acc act gtt gtg gga gag ccc ate acc ate ccc aag 1244 Ser Lys Pro lie Thr Thr Val Val Gly Glu Pro lie Thr lie Pro Lys 165 170 175 ctg gag cac cca acc cag caa gac ate gac ctg tac cac acc atg tac 1292 Leu Glu His Pro Thr Gin Gin Asp lie Asp Leu Tyr His Thr Met Tyr 180 185 190 atg gag gcc ctg gtg aag ctc ttc gac aag cac aag acc aag ttc ggc 1340 Met Glu Ala Leu Val Lys Leu Phe Asp Lys His Lys Thr Lys Phe Gly 195 200 205 ctc ccg gag act gag gtc ctg gag gtg aac tga gccagccttc ggggccaatt 1393 Leu Pro Glu Thr Glu Val Leu Glu Val Asn 210 215 WO 02/083721 - 8 - PCT/SE02/00730 ccctggagga accagctgca aatcactttt ttgctctgta aatttggaag tgtcatgggt 1453 gtctgtgggt tatttaaaag aaattataac aattttgcta aaccattaca atgttaggtc 1513 ttttttaaga aggaaaaagt cagtatttca agttctttca cttccagctt gccctgttct 1573 aggtggtggc taaatctggg cctaatctgg gtggctcagc taacctctct tcttcccttc 1633 ctgaagtgac aaaggaaact cagtcttctt ggggaagaag gattgccatt agtgacttgg 1693 accagttaga tgattcactt tttgccccta gggatgagag gcgaaagcca cttctcatac 1753 aagccccttt attgccacta ccccacgctc gtctagtcct gaaactgcag gaccagtttc 1813 tctgccaagg ggaggagttg gagagcacag ttgccccgtt gtgtgagggc agtagtaggc 1873 atctggaatg ctccagtttg atctcccttc tgccacccct acctcacccc tagtcactca 1933 tatcggagcc tggactggcc tccaggatga ggatgggggt ggcaatgaca ccctgcaggg 1993 gaaaggactg ccccccatgc accattgcag ggaggatgcc gccaccatga gctaggtgga 2053 gtaactggtt tttcttgggt ggctgatgac atggatgcag cacagactca gccttggcct 2113 ggagcacatg cttactggtg gcctcagttt accttcccca gatcctagat tctggatgtg 2173 aggaagagat ccctcttcag aaggggcctg gccttctgag cagcagatta gttccaaagc 2233 aggtggcccc cgaacccaag cctcactttt ctgtgccttc ctgagggggt tgggccgggg 2293 aggaaaccca accctctcct gtgtgttctg ttatctcttg atgagatcat tgcaccatgt 2353 cagacttttg tatatgcctt gaaaataaat gaaagtgaga catggtgcaa tgatctcatc 2413 aagagataac agaacagaca ggagagggtt gggttatctc ttgatgagat cattgcacca 2473 tgtcagactt ttgtatatgc cttgaaaata aatgaaagtg agaatc 2519 <210> 8 <211> 219 <212> PRT <213> human <400> 8 Ala Phe Cys Asn Phe Ser Thr Glu Ala Thr Glu Val Ser Lys Lys Phe 1 5 10 15 Pro Gly lie Arg Pro Tyr Leu Ala Thr Leu Ala Gly Asn Phe Arg Met 20 25 30 Pro Val Leu Arg Glu Tyr Leu Met Ser Gly Gly lie Cys Pro Val Ser 35 40 45 Arg Asp Thr lie Asp Tyr Leu Leu Ser Lys Asn Gly Ser Gly Asn Ala 50 55 60 lie lie lie Val Val Gly Gly Ala Ala Glu Ser Leu Ser Ser Met Pro 65 70 75 80 WO 02/083721 -9- PCT/SE02/00730 Gly Lys Asn Ala Val Thr Leu Arg Asn 85 Ala Leu Arg His Gly Ala Asp Leu Val 100 105 Asn Glu Val Tyr Lys Gin Val lie Phe 115 120 Trp Val Gin Lys Lys Phe Gin Lys Tyr 130 135 Phe His Gly Arg Gly Leu Phe Ser Ser 145 150 Tyr Ser Lys Pro lie Thr Thr Val Val 165 Lys Leu Glu His Pro Thr Gin Gin Asp 180 185 Tyr Met Glu Ala Leu Val Lys Leu Phe 195 200 Gly Leu Pro Glu Thr Glu Val Leu Glu 210 215 <210> 9 <211> 685 <212> DNA <213> human <220> <221> misc_feature <222> ()..() <223> n = A, C. G or T <220> <221> CDS <222> (2).. (322) <400> 9 g egg aac cgc aag ggc ttt gtg aaa ctg gcc ctg cgt cat gga get gac 4 9 Arg Asn Arg Lys Gly Phe Val Lys Leu Ala Leu Arg His Gly Ala Asp 15 10 15 ctg gtt ccc ate tac tcc ttt gga gag aat gaa gtg tac aag cag gtg 97 Leu Val Pro lie Tyr Ser Phe Gly Glu Asn Glu Val Tyr Lys Gin Val 20 25 30 ate ttc gag gag ggc tcc tgg ggc cga tgg gtc cag aag aag ttc cag 145 lie Phe Glu Glu Gly Ser Trp Gly Arg Trp Val Gin Lys Lys Phe Gin 35 40 45 aaa tac att ggt ttc gcc cca tgc ate ttc cat ggt cga ggc ctc ttc 193 Lys Tyr lie Gly Phe Ala Pro Cys lie Phe His Gly Arg Gly Leu Phe 50 55 60 Arg Lys Gly Phe Val Lys Leu 90 95 Pro lie Tyr Ser Phe Gly Glu 110 Glu Glu Gly Ser Trp Gly Arg 125 lie Gly Phe Ala Pro Cys lie 140 Asp Thr Trp Gly Leu Val Pro 155 160 Gly Glu Pro lie Thr lie Pro 170 175 lie Asp Leu Tyr His Thr Met 190 Asp Lys His Lys Thr Lys Phe 205 Val Asn WO 02/083721 PCT/SE02/00730 -10- tcc tcc gac acc tgg ggg ctg gtg ccc tac tcc aag ccc ate acc act 241 Ser Ser Asp Thr Trp Gly Leu Val Pro Tyr Ser Lys Pro lie Thr Thr 65 70 75 80 gtt ggt ggt gga aaa att cag tct agg agt aaa aaa agg aag ate aac 289 Val Gly Gly Gly Lys lie Gin Ser Arg Ser Lys Lys Arg Lys lie Asn 85 90 95 atn ntg cag aat gac tea tgc tat tea tta tag aagcaattgc tggagatgnt 342 Xaa Xaa Gin Asn Asp Ser Cys Tyr Ser Leu 100 105 atcattgtgg ateaeggaag tcttcatgga agaggtggca tttgagctgg gccttcactg 4 02 aagcggtgaa tcggcgtcct gggtgcctgg cacaccttgt agctcagctt actagctagt 4 62 ggagtgcgaa ggggcgtgta cttgtcggtt ggagctggtc atgaaagagc tcgtgggact 522 gcccgacggt tctcaggtcc cagtgcatcc tgcgtggtgg ctctctgctg aaccataaag 582 cattcctttt caatccctgc acgctcacgc cgggaaaaga ctgcacaagg ggctccaagg 642 cagacaagcg atcgccaccc agctggcttc cgagggtccc cgc 685 <210> 10 <211> 106 <212> PRT <213> human <220> <221> misc_feature <222> {)..() <223> n = A, C. G or T <4 00> 10 Arg Asn Arg Lys Gly Phe Val Lys Leu Ala Leu Arg His Gly Ala Asp 15 10 15 Leu Val Pro lie Tyr Ser Phe Gly Glu Asn Glu Val Tyr Lys Gin Val 20 25 30 lie Phe Glu Glu Gly Ser Trp Gly Arg Trp Val Gin Lys Lys Phe Gin 35 40 45 Lys Tyr lie Gly Phe Ala Pro Cys lie Phe His Gly Arg Gly Leu Phe 50 55 60 Ser Ser Asp Thr Trp Gly Leu Val Pro Tyr Ser Lys Pro lie Thr Thr 65 70 75 80 Val Gly Gly Gly Lys lie Gin Ser Arg Ser Lys Lys Arg Lys lie Asn 85 90 95 Xaa Xaa Gin Asn Asp Ser Cys Tyr Ser Leu 100 105 <210> 11 <211> 474 WO 02/083721 -11 - PCT/SE02/00730 <212> DNA <213> human <220> <221> CDS < 2 2 2 > (2).. (442) <400> 11 a aaa gaa tea ctg gat get cat cct gga aag ttc act ctg ttc ate cgc 4 9 Lys Glu Ser Leu Asp Ala His Pro Gly Lys Phe Thr Leu Phe lie Arg 15 10 15 cag egg aaa gga ttt gtt aaa att get ttg acc cat ggc gcc tct ctg 97 Gin Arg Lys Gly Phe Val Lys lie Ala Leu Thr His Gly Ala Ser Leu 20 25 30 gtc cca gtg gtt tct ttt ggt gaa aat gaa ctg ttt aaa caa act gac 145 Val Pro Val Val Ser Phe Gly Glu Asn Glu Leu Phe Lys Gin Thr Asp 35 40 45 aac cct gaa gga tea tgg att aga act gtt cag aat aaa ctg cag aag 193 Asn Pro Glu Gly Ser Trp lie Arg Thr Val Gin Asn Lys Leu Gin Lys 50 55 - 60 ate atg ggg ttt get ttg ccc ctg ttt cat gcc agg gga gtt ttt cag 241 lie Met Gly Phe Ala Leu Pro Leu Phe His Ala Arg Gly Val Phe Gin 65 70 75 80 tac aat ttt ggc eta atg acc tat agg aaa gcc ate cac act gtt gtt 289 Tyr Asn Phe Gly Leu Met Thr Tyr Arg Lys Ala lie His Thr Val Val 85 90 95 ggc cgc ccg ate cct gtt cgt cag act ctg aac ccg acc cag gag cag 337 Gly Arg Pro lie Pro Val Arg Gin Thr Leu Asn Pro Thr Gin Glu Gin 100 105 110 att gag gag tta cat cag acc tat atg gag gaa ctt agg aaa ttg ttt 385 lie Glu Glu Leu His Gin Thr Tyr Met Glu Glu Leu Arg Lys Leu Phe 115 120 125 gag gaa cac aaa gga aag tat ggc att cca gag cac gag act ctt gtt 433 Glu Glu His Lys Gly Lys Tyr Gly lie Pro Glu His Glu Thr Leu Val 130 135 140 tta aaa tga cttgactata aaaaaaaaaa aaaageggee gc 474 Leu Lys 145 <210> 12 <211> 146 <212> PRT <213> human <4 00> 12 Lys Glu Ser Leu Asp Ala His Pro Gly Lys Phe Thr Leu Phe lie Arg 15 10 15 Gin Arg Lys Gly Phe Val Lys lie Ala Leu Thr His Gly Ala Ser Leu 20 25 30 WO 02/083721 -12- PCT/SE02/00730 Val Pro Val Val Ser Phe Gly Glu Asn Glu Leu Phe Lys Gin Thr Asp 35 40 45 Asn Pro Glu Gly Ser Trp lie Arg Thr Val G'ln Asn Lys Leu Gin Lys 50 55 60 lie Met Gly Phe Ala Leu Pro Leu Phe His Ala Arg Gly Val Phe Gin 65 70 75 80 Tyr Asn Phe Gly Leu Met. Thr Tyr Arg Lys Ala lie His Thr Val Val 85 90 95 Gly Arg Pro lie Pro Val Arg Gin Thr Leu Asn Pro Thr Gin Glu Gin 100 105 110 lie Glu Glu Leu His Gin Thr Tyr Met Glu Glu Leu Arg Lys Leu Phe 115 120 125 Glu Glu His Lys Gly Lys Tyr Gly lie Pro Glu His Glu Thr Leu Val 130 135 140 Leu Lys 145 <210> 13 <211> 254 <212> DNA <213> human <220> <221> misc ; <222> 0 • • ( <223> n = A, <220> <221> CDS <222> (3) . . <400> 13 gc aac ctc ttc gag gcc cac aaa ctt aag ttc aac ate att gta ggg Asn Leu Phe Glu Ala His Lys Leu Lys Phe Asn lie lie Val Gly 15 10 15 47 ggt gcc cag gag gcc ctg gat gcc agg cct gga tcc ttc acg ctg tta Gly Ala Gin Glu Ala Leu Asp Ala Arg Pro Gly Ser Phe Thr Leu Leu 20 25 30 95 ctg egg aac cga aag ggc ttc gtc agg ctc gcc ctg aca cac ggg gca Leu Arg Asn Arg Lys Gly Phe Val Arg Leu Ala Leu Thr His Gly Ala 35 40 45 143 ccc ctg gtt nta ate ttc tcc ttc ggg gag aat gac eta ttt gac cag 191 Pro Leu Val Xaa lie Phe Ser Phe Gly Glu Asn Asp Leu Phe Asp Gin 50 55 60 att ccc aac tct tct ggc tcc tgg tta cgc tat ate cag aat egg ttg 239 lie Pro Asn Ser Ser Gly Ser Trp Leu Arg Tyr lie Gin Asn Arg Leu 65 70 75 WO 02/083721 - 13- PCT/SE02/00730 cag aag ate atg ggc 254 Gin Lys lie Met Gly 80 <210> 14 <211> 84 <212> PRT <213> human <220> <221> misc_feature <222> ()..() <223> n = A, C, G or T <400> 14 Asn Leu Phe Glu Ala His Lys Leu Lys Phe Asn lie lie Val Gly Gly 15 10 15 Ala Gin Glu Ala Leu Asp Ala Arg Pro Gly Ser Phe Thr Leu Leu Leu 20 25 30 Arg Asn Arg Lys Gly Phe Val Arg Leu Ala Leu Thr His Gly Ala Pro 35 40 45 Leu Val Xaa lie Phe Ser Phe Gly Glu Asn Asp Leu Phe Asp Gin lie 50 55 60 Pro Asn Ser Ser Gly Ser Trp Leu Arg Tyr lie Gin Asn Arg Leu Gin 65 70 75 80 Lys lie Met Gly <210> 15 <211> 887 <212> DNA <213> human <220> <221> CDS <222> (314)..(805) <400> 15 ggctgtttca gcatggcggt gcctccatgt ggccttttgg tgtcttcatg ttatatcctg 60 tccaggtggt gttggtataa ataattctag gcaccatcat acctgagttt ctcagtagcc 120 ctaggaggta gcagggacag gtccaaatac tctattgcca ctttacaaat gaagagcctg 180 taggagaggg aagcaatttg tcccaagcca gcatcaagtc tgtggcacag ccagcaccat 240 aatatctcca ggtgctgtca cataccatat ctgaatcttc gtaagaaccc agggtggtca 300 gacatatgga tga aga cct gga ggc tea gag ggg agg ttt ccc aag gtc 349 Arg Pro Gly Gly Ser Glu Gly Arg Phe Pro Lys Val 15 10 WO 02/083721 - 14- PCT/SE02/00730 aca cca gtg agt ggc aga gtc agg get ggt aca cag gcc ccg ccc tgg 397 Thr Pro Val Ser Gly Arg Val Arg Ala Gly Thr Gin Ala Pro Pro Trp 15 20 25 ctc age agg ttg ccg tcc ctg cag ctg gtc aag act get gag ctg gac 44 5 Leu Ser Arg Leu Pro Ser Leu Gin Leu Val Lys Thr Ala Glu Leu Asp 30 35 40 ccc tct egg aac tac att gcg ggc ttc cac ccc cat gga gtc ctg gca 4 93 Pro Ser Arg Asn Tyr lie Ala Gly Phe His Pro His Gly Val Leu Ala 45 50 55 60 gtc gga -gcc ttt gcc aac ctg tgc act gag age aca ggc ttc tct teg 541 Val Gly Ala Phe Ala Asn Leu Cys Thr Glu Ser Thr Gly Phe Ser Ser 65 70 75 ate ttc ccc ggt ate cgc ccc cat ctg atg atg ctg acc ttg tgg ttc 589 lie Phe Pro Gly lie Arg Pro His Leu Met Met Leu Thr Leu Trp Phe 80 85 90 egg gcc ccc ttc ttc aga gat tac ate atg tct gca ggg ttg gtc aca 637 Arg Ala Pro Phe Phe Arg Asp Tyr lie Met Ser Ala Gly Leu Val Thr 95 100 105 tea gaa aag gag agt get get cac att ctg aac agg aag ggt ggc gga 685 Ser Glu Lys Glu Ser Ala Ala His lie Leu Asn Arg Lys Gly Gly Gly 110 115 120 aac ttg ctg ggc ate att gta ggg ggt gcc cag gag gcc ctg gat gcc 733 Asn Leu Leu Gly lie lie Val Gly Gly Ala Gin Glu Ala Leu Asp Ala 125 130 135 140 agg cct gga tcc ttc acg ctg tta ctg egg aac cga aag ggc ttc gtc 781 Arg Pro Gly Ser Phe Thr Leu Leu Leu Arg Asn Arg Lys Gly Phe Val 145 150 155 agg ctc gcc ctg aca cac ggg tat caagcctctg ggaagagcac tctgggttca 835 Arg Leu Ala Leu Thr His Gly Tyr 160 gttggcaatt ggcaagcgat ctttattttg gtgggaagat ggcagagacg aa 887 <210> 16 <211> 164 <212> PRT <213> human <400> 16 Arg Pro Gly Gly Ser Glu Gly Arg Phe Pro Lys Val Thr Pro Val Ser 15 10 15 Gly Arg Val Arg Ala Gly Thr Gin Ala Pro Pro Trp Leu Ser Arg Leu 20 25 30 Pro Ser Leu Gin Leu Val Lys Thr Ala Glu Leu Asp Pro Ser Arg Asn 35 40 45 Tyr lie Ala Gly Phe His Pro His Gly Val Leu Ala Val Gly Ala Phe 50 55 60 WO 02/083721 - 15- PCT/SE02/00730 Ala Asn Leu Cys Thr Glu Ser Thr 65 70 lie Arg Pro His Leu Met Met Leu 85 Phe Arg Asp Tyr lie Met Ser Ala 100 Ser Ala Ala His lie Leu Asn Arg 115 120 He lie Val Gly Gly Ala Gin Glu 130 135 Phe Thr Leu Leu Leu Arg Asn Arg 14 5 150 Thr His Gly Tyr Gly Phe Ser Ser lie Phe Pro Gly 75 80 Thr Leu Trp Phe Arg Ala Pro Phe 90 95 Gly Leu Val Thr Ser Glu Lys Glu 105 110 Lys Gly Gly Gly Asn Leu Leu Gly 125 Ala Leu Asp Ala Arg Pro Gly Ser 140 Lys Gly Phe Val Arg Leu Ala Leu 155 160 <210> 17 <211> 446 <212> DNA <213> human <220> <221> CDS <222> (1).. (444 ) <400> 17 age gat tat gtc cct ctc aag ctt ctg aag act cat gac ate tgc ccc 4 8 Ser Asp Tyr Val Pro Leu Lys Leu Leu Lys Thr His Asp lie Cys Pro 15 10 15 age cgc aac tac ate ctc gtc tgc cac cct cat ggg ctc ttt gcc cat 96 Ser Arg Asn Tyr lie Leu Val Cys His Pro His Gly Leu Phe Ala His 20 25 30 gga tgg ttt ggc cac ttt gcc aca gag gcc tea ggc ttc tcc aag ata 14 4 Gly Trp Phe Gly His Phe Ala Thr Glu Ala Ser Gly Phe Ser Lys lie 35 40 45 ttt cct ggc ate acc cct tac ata ctc aca ctg gga gcc ttt ttc tgg 192 Phe Pro Gly lie Thr Pro Tyr lie Leu Thr Leu Gly Ala Phe Phe Trp 50 55 60 atg cct ttc ctc aga gaa tat gta atg tct aca ggg gcc tgc tct gtg 240 Met Pro Phe Leu Arg Glu Tyr Val Met Ser Thr Gly Ala Cys Ser Val 65 70 75 80 agt cga tcc tcc att gac ttt ctg ctg act cat aaa ggc aca ggc aac 288 Ser Arg Ser Ser lie Asp Phe Leu Leu Thr His Lys Gly Thr Gly Asn 85 90 95 atg gtc att gtg gtg att ggt gga ctg get gag tgc aga tac age ctg 336 Met Val lie Val Val lie Gly Gly Leu Ala Glu Cys Arg Tyr Ser Leu 100 105 110 WO 02/083721 - 16- PCT/SE02/00730 cca ggt tct tct acc ctg Pro Gly Ser Ser Thr Leu 115 atg gcc ctt cag cat ggg Met Ala Leu Gin His Gly 130 gag acg gac ctc ta Glu Thr Asp Leu 145 gtg ttg aag aac egg tct Val Leu Lys Asn Arg Ser 120 gtg cct eta ata cct gcc Val Pro Leu lie Pro Ala 135 140 ggc ttt gtg cgc 384 Gly Phe Val Arg 125 tat gcc ttt ggg 432 Tyr Ala Phe Gly 446 <210> 18 <211> 148 <212> PRT <213> human <400> 18 Ser Asp Tyr Val Pro Leu Lys Leu Leu Lys Thr His Asp lie Cys Pro 15 10 15 Ser Arg Asn Tyr lie Leu Val Cys His Pro His Gly Leu Phe Ala His 20 25 30 Gly Trp Phe Gly His Phe Ala Thr Glu Ala Ser Gly Phe Ser Lys lie 35 40 45 Phe Pro Gly lie Thr Pro Tyr lie Leu Thr Leu Gly Ala Phe Phe Trp 50 55 60 Met Pro Phe Leu Arg Glu Tyr Val Met Ser Thr Gly Ala Cys Ser Val 65 70 75 80 Ser Arg Ser Ser lie Asp Phe Leu Leu Thr His Lys Gly Thr Gly Asn 85 90 95 Met Val lie Val Val lie Gly Gly Leu Ala Glu Cys Arg Tyr Ser Leu 100 105 110 Pro Gly Ser Ser Thr Leu Val Leu Lys Asn Arg Ser Gly Phe Val Arg 115 120 125 Met Ala Leu Gin His Gly Val Pro Leu lie Pro Ala Tyr Ala Phe Gly 130 135 140 Glu Thr Asp Leu 145 <210> 19 <211> 1670 <212> DNA <213> human <220> <221> misc_feature <222> ()..() WO 02/083721 -17- PCT/SE02/00730 <223> n = A, C, G or T <220> <221> CDS <222> (635) . . (1666) <4 00> 19 gggaagagaa tatcgttttt ettgeaaaat acacgctaaa aactatttag aagcaaaagg 60 ttgtaatctc tgtgatgtat tctcaaatac aaacatatat gtatatactt acatttttac 120 atttaaagat aaatcaaacg taaaatgttg acaatgggta gatgtagatg aagattaaac 180 aagactttat taaaataatc ttgttttttc aaaataaaaa gtttaattaa aaaacctcca 240 tcaagagttt ttgtagcaat aaacaagctg attcaaaaat ttatatagaa aaacaaagaa 300 actacaaata attaaaacaa ttttgagaac gaataaagtt aaaggaatta taccatctga 360 ttttgagact tagcataaga ctagagcaat caagacagtg atgtatttgt gaaggaatag 420 atatattgat ccacagaaca gaaaagagtc aagaaataaa cacatgaata tggtcaattg 480 atttttgaca aagatgaaaa agcaattcca tggaggatga ataagtgett ttcaaggaac 540 ggtgtaggaa aatttgatgt ccatatgtgg caaaatgaat cttgacccaa acttcaggct 600 ctataaaaat taactcaagt atgacatcaa caag atg gtg aat ggg aag tcc ate 655 Met Val Asn Gly Lys Ser He 1 5 aca tct ctc cag age aac aag aat ctg gca gcc ate cat gga cca aag 703 Thr Ser Leu Gin Ser Asn Lys Asn Leu Ala Ala lie His Gly Pro Lys 10 15 20 tac ctt tgt ggg aat ttt gga ccc agg tgg cag gcg ttc age ttg ggt 7 51 Tyr Leu Cys Gly Asn Phe Gly Pro Arg Trp Gin Ala Phe Ser Leu Gly 25 30 35 acg aaa ctg gac cct atg gaa gta ttt ccg aaa tta ctt ccc agt aaa 799 Thr Lys Leu Asp Pro Met Glu Val Phe Pro Lys Leu Leu Pro Ser Lys 40 45 50 55 gtc cct gtt gcc cag acc ctt get ccc tac tea get cca tgt ttt cag 847 Val Pro Val Ala Gin Thr Leu Ala Pro Tyr Ser Ala Pro Cys Phe Gin 60 65 70 agg ctt tgg tgg tea gca gcg aag gtc aag gcc ccg agt cat aat gca 8 95 Arg Leu Trp Trp Ser Ala Ala Lys Val Lys Ala Pro Ser His Asn Ala 75 80 85 aag caa ggg ccc aag atg gat ggg cag ctg gtg aag act cat gat ctt 94 3 Lys Gin Gly Pro Lys Met Asp Gly Gin Leu Val Lys Thr His Asp Leu 90 95 100 tct ccc aaa cac aac tac ate att gcc aat cac ccc cat ggc att ctc 991 Ser Pro Lys His Asn Tyr lie lie Ala Asn His Pro His Gly lie Leu 105 110 115 WO 02/083721 - 18- PCT/SE02/00730 tct ttt ggt gtc ttc ate aac ttt gcc act gag gcc act ggc att get 1039 Ser Phe Gly Val Phe lie Asn Phe Ala Thr Glu Ala Thr Gly lie Ala 120 125 130 135 egg att ttc cca tcc ate act ccc ttt gta ggg acc tta gaa agg ata 1087 Arg lie Phe Pro Ser lie Thr Pro Phe Val Gly Thr Leu Glu Arg lie 140 145 150 ttt tgg ate cca att gtg cga gaa tat gtg atg tea atg ggt gtg tgc 1135 Phe Trp lie Pro lie Val Arg Glu Tyr Val Met Ser Met Gly Val Cys 155 160 165 cct gtg agt age tea gcc ttg aag tac ttg ctg acc cag aaa ggc tea 1183 Pro Val Ser Ser Ser Ala Leu Lys Tyr Leu Leu Thr Gin Lys Gly Ser 170 175 180 ggc aat gcc gtg gtt att gtg gtg ggt gga get get gaa get ctc ttg 1231 Gly Asn Ala Val Val lie Val Val Gly Gly Ala Ala Glu Ala Leu Leu 185 190 195 tgc cga cca gga gcc tcc act ctc ttc ctc aag cag cgt aaa ggt ttt 1279 Cys Arg Pro Gly Ala Ser Thr Leu Phe Leu Lys Gin Arg Lys Gly Phe 200 205 210 215 gtg aag atg gca ctg caa aca ggg gca tac ctt gtc cct tea tat tcc 1327 Val Lys Met Ala Leu Gin Thr Gly Ala Tyr Leu Val Pro Ser Tyr Ser 220 225 230 ttt ggt gag aac gaa gtt ttc aat cag gag acc ttc cct gag ggc acg 1375 Phe Gly Glu Asn Glu Val Phe Asn Gin Glu Thr Phe Pro Glu Gly Thr 235 240 245 tgg tta agg ttg ttc caa aaa acc ttc cag gac aca ttc aaa aaa ate 1423 Trp Leu Arg Leu Phe Gin Lys Thr Phe Gin Asp Thr Phe Lys Lys lie 250 255 260 ctg gga eta aat ttc tgt acc ttc cat ggc egg ggc ttc act cgc gga 1471 Leu Gly Leu Asn Phe Cys Thr Phe His Gly Arg Gly Phe Thr Arg Gly 265 270 275 tcc tgg ggc ttc ctg cct ttc aat egg ccc att acc act gtt gtt ggg 1519 Ser Trp Gly Phe Leu Pro Phe Asn Arg Pro lie Thr Thr Val Val Gly 280 285 290 295 gaa ccc ctt cca att ccc agg att aag agg cca aac cag aag aca gta 1567 Glu Pro Leu Pro lie Pro Arg lie Lys Arg Pro Asn Gin Lys Thr Val 300 305 310 gac aag tat cac gca ctc tac ate agt gcc ctg cgc aag ctc ttt gac 1615 Asp Lys Tyr His Ala Leu Tyr lie Ser Ala Leu Arg Lys Leu Phe Asp 315 320 325 caa cac aaa gtt gaa tat ggc ctc cct gag acc caa gag ctg aca att 1663 Gin His Lys Val Glu Tyr Gly Leu Pro Glu Thr Gin Glu Leu Thr lie 330 335 340 aca ntaa Thr 1670 WO 02/083721 -19- PCT/SE02/00730 <210> 20 <211> 344 <212> PRT <213> human <220> <221> misc_feature <222> ()..() <223> n = A, C, G or T <400> 20 Met Val Asn Gly Lys Ser lie Thr Ser Leu Gin Ser Asn Lys Asn Leu 15 10 15 Ala Ala lie His Gly Pro Lys Tyr Leu Cys Gly Asn Phe Gly Pro Arg 20 25 30 Trp Gin Ala Phe Ser Leu Gly Thr Lys Leu Asp Pro Met Glu Val Phe 35 40 45 Pro Lys Leu Leu Pro Ser Lys Val Pro Val Ala Gin Thr Leu Ala Pro 50 55 60 Tyr Ser Ala Pro Cys Phe Gin Arg Leu Trp Trp Ser Ala Ala Lys Val 65 70 75 80 Lys Ala Pro Ser His Asn Ala Lys Gin Gly Pro Lys Met Asp Gly Gin 85 90 95 Leu Val Lys Thr His Asp Leu Ser Pro Lys His Asn Tyr lie lie Ala 100 105 110 Asn His Pro His Gly lie Leu Ser Phe Gly Val Phe lie Asn Phe Ala 115 120 125 Thr Glu Ala Thr Gly lie Ala Arg lie Phe Pro Ser lie Thr Pro Phe 130 135 140 Val Gly Thr Leu Glu Arg lie Phe Trp lie Pro lie Val Arg Glu Tyr 145 150 155 160 Val Met Ser Met Gly Val Cys Pro Val Ser Ser Ser Ala Leu Lys Tyr 165 170 175 Leu Leu Thr Gin Lys Gly Ser Gly Asn Ala Val Val lie Val Val Gly 180 185 190 Gly Ala Ala Glu Ala Leu Leu Cys Arg Pro Gly Ala Ser Thr Leu Phe 195 200 205 Leu Lys Gin Arg Lys Gly Phe Val Lys Met Ala Leu Gin Thr Gly Ala 210 215 220 Tyr Leu Val Pro Ser Tyr Ser Phe Gly Glu Asn Glu Val Phe Asn Gin 225 230 235 240 Glu Thr Phe Pro Glu Gly Thr Trp Leu Arg Leu Phe Gin Lys Thr Phe 245 250 255 WO 02/083721 -20- PCT/SE02/00730 Gin Asp Thr Phe Lys Lys lie Leu 260 Gly Arg Gly Phe Thr Arg Gly Ser 275 280 Pro lie Thr Thr Val Val Gly Glu 290 295 Arg Pro Asn Gin Lys Thr Val Asp 305 310 Ala Leu Arg Lys Leu Phe Asp Gin 325 Glu Thr Gin Glu Leu Thr lie Thr 340 Gly Leu Asn Phe Cys Thr Phe His 265 270 Trp Gly Phe Leu Pro Phe Asn Arg 285 Pro Leu Pro lie Pro Arg lie Lys 300 Lys Tyr His Ala Leu Tyr lie Ser 315 320 His Lys Val Glu Tyr Gly Leu Pro 330 335