WO1999047667A1 - Human sec61 gamma subunit gene: cbdame07 - Google Patents

Abstract

CBDAME07 polypeptides and polynucleotides and methods for producing such polypeptides by recombinant techniques are disclosed. Also disclosed are methods for utilizing CBDAME07 polypeptides and polynucleotides in the design of protocols for the treatment of cancer, AIDS, metabolic disorders, and IDDM, among others, and diagnostic assays for such conditions.

Description

Human Sec61 Gamma Subunit Gene: CBDAME07

FIELD OF INVENTION

This invention relates to newly identified polynucleotides, polypeptides encoded by them and to the use of such polynucleotides and polypeptides, and to their production. More particularly, the polynucleotides and polypeptides of the present invention relate to the protein transporter sec61 family, hereinafter referred to as CBDAME07 . The invention also relates to inhibiting or activating the action of such polynucleotides and polypeptides.

BACKGROUND OF THE INVENTION

The protein transporter sec61 family is highly conserved in organisms including C. elegans, yeast, rice to mammals. This indicates that the protein transporter secόl family has an established, proven history as therapeutic targets. Clearly there is a need for identification and characterization of further members of the protein transporter sec61 family which can play a role in preventing, ameliorating or correcting dysfunctions or diseases, including, but not limited to, cancer, AIDS, metabolic disorders, and insulin dependent diabetes mellitus (IDDM).

SUMMARY OF THE INVENTION In one aspect, the invention relates to CBDAME07 polypeptides and recombinant materials and methods for their production. Another aspect of the invention relates to methods for using such CBDAME07 polypeptides and polynucleotides. Such uses include the treatment of cancer, AIDS, metabolic disorders, and IDDM, among others. In still another aspect, the invention relates to methods to identify agonists and antagonists using the materials provided by the invention, and treating conditions associated with CBDAME07 imbalance with the identified compounds. Yet another aspect of the invention relates to diagnostic assays for detecting diseases associated with inappropriate CBDAME07 activity or levels.

DESCRIPTION OF THE INVENTION Definitions

The following definitions are provided to facilitate understanding of certain terms used frequently herein.

"CBDAME07 " refers, among others, generally to a polypeptide having the amino acid sequence set forth in SEQ ID NOJ or an allelic variant thereof.

1 "CBDAME07 activity or CBDAME07 polypeptide activity" or "biological activity of the CBDAME07 or CBDAME07 polypeptide" refers to the metabolic or physiologic function of said CBDAME07 including similar activities or improved activities or these activities with decreased undesirable side-effects Also included are antigenic and lmmunogenic activities of said CBDAME07

"CBDAME07 gene" refers to a polynucleotide having the nucleotide sequence set forth in SEQ ID NO 1 or alle c vanants thereof and/or their complements

"Antibodies" as used herem includes polyclonal and monoclonal antibodies, chimenc, smgle chain, and humamzed antibodies, as well as Fab fragments, including the products of an Fab or other lmmunoglobulin expression library

"Isolated" means altered "by the hand of man" from the natural state If an "isolated" composition or substance occurs in nature, it has been changed or removed from its original environment, or both For example, a polynucleotide or a polypeptide naturally present in a living animal is not "isolated ' but the same polynucleotide or polypeptide separated from the coexisting materials of its natural state is "isolated", as the term is employed herein

"Polynucleotide" generally refers to any polynbonucleotide or polydeoxπbonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA "Polynucleotides" include, without limitation smgle- and double-stranded DNA, DNA that is a mixture of smgle- and double- stranded regions, smgle- and double-stranded RNA, and RNA that is mixture of smgle- and double-stranded regions, hybnd molecules compnsing DNA and RNA that may be smgle-stranded or, more typically, double-stranded or a mixture of smgle- and double-stranded regions In addition, "polynucleotide" refers to tnple-stranded regions comprising RNA or DNA or both RNA and DNA The term polynucleotide also mcludes DNAs or RNAs contammg one or more modified bases and DNAs or RNAs with backbones modified for stability or for other reasons "Modified" bases include, for example, tntylated bases and unusual bases such as mosine A variety of modifications has been made to DNA and RNA, thus, "polynucleotide" embraces chemically, enzymatically or metabohcally modified forms of polynucleotides as typically found m nature, as well as the chemical forms of DNA and RNA charactenstic of viruses and cells "Polynucleotide" also embraces relatively short polynucleotides, often referred to as ohgonucleotides "Polypeptide" refers to any peptide or protein comprising two or more ammo acids joined to each other by peptide bonds or modified peptide bonds, l e , peptide isosteres "Polypeptide" refers to both short chains, commonly referred to as peptides, ohgopeptides or ohgomers, and to longer chains, generally referred to as proteins Polypeptides may contain ammo acids other than the 20 gene-encoded ammo acids "Polypeptides" include ammo acid sequences modified either by

2 natural processes, such as posttranslational processing, or by chemical modification techniques which are well known in the art. Such modifications are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature. Modifications can occur anywhere in a polypeptide, including the peptide backbone, the amino acid side-chains and the amino or carboxyl termini. It will be appreciated that the same type of modification may be present in the same or varying degrees at several sites in a given polypeptide. Also, a given polypeptide may contain many types of modifications. Polypeptides may be branched as a result of ubiquitination, and they may be cyclic, with or without branching. Cyclic, branched and branched cyclic polypeptides may result from posttranslation natural processes or may be made by synthetic methods. Modifications include acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cystine, formation of pyroglutamate, formylation, gamma- carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to proteins such as arginylation, and ubiquitination. See, for instance, PROTEINS - STRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, New York, 1993 and Wold, F., Posttranslational Protein Modifications: Perspectives and Prospects, pgs. 1-12 in POSTTRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, B. C. Johnson, Ed., Academic Press, New York, 1983; Seifter et al., "Analysis for protein modifications and nonprotein cofactors", Meth Enzymol (1990) 182:626-646 and Rattan et al, "Protein Synthesis: Posttranslational Modifications and Aging", Ann NY Acad Sci (1992) 663:48-62. "Variant" as the term is used herein, is a polynucleotide or polypeptide that differs from a reference polynucleotide or polypeptide respectively, but retains essential properties. A typical variant of a polynucleotide differs in nucleotide sequence from another, reference polynucleotide. Changes in the nucleotide sequence of the variant may or may not alter the amino acid sequence of a polypeptide encoded by the reference polynucleotide. Nucleotide changes may result in amino acid substitutions, additions, deletions, fusions and truncations in the polypeptide encoded by the reference sequence, as discussed below. A typical variant of a polypeptide differs in amino acid sequence from another, reference polypeptide. Generally, differences are limited so that the sequences of the reference polypeptide and the variant are closely similar overall and, in many regions, identical. A variant and reference polypeptide may differ in amino acid sequence by one or

3 more substitutions, additions, deletions m any combination A substituted or inserted amino acid residue may or may not be one encoded by the genetic code A variant of a polynucleotide or polypeptide may be a naturally occurring such as an allehc vanant, or it may be a vanant that is not known to occur naturally Non-naturally occurring vanants of polynucleotides and polypeptides may be made by mutagenesis techniques or by direct synthesis

"Identity," as known m the art, is a relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as determined by companng the sequences In the art, "identity" also means the degree of sequence relatedness between polypeptide or polynucleotide sequences, as the case may be, as determined by the match between strings of such sequences "Identity" and "similarity" can be readily calculated by known methods, including but not limited to those described in (Computational Molecular Biology, Lesk, A M , ed , Oxford University Press, New York, 1988, Biocomputmg Informatics and Genome Projects, Smith, D W , ed , Academic Press, New York, 1993, Computer Analysis of Sequence Data, Part I, Gnffin, A M , and Gnffin, H G , eds , Humana Press, New Jersey, 1994, Sequence Analysis in Molecular Biology, von Hemje, G , Academic Press, 1987, and Sequence Analysis Pnmer, Gnbskov, M and Devereux, J , eds , M Stockton Press, New York, 1991, and Canllo, H , and Lipman, D , SLAM J Applied Math , 48 1073 (1988) Preferred methods to determme identity are designed to give the largest match between the sequences tested Methods to determme identity and similarity are codified in publicly available computer programs Preferred computer program methods to determme identity and similanty between two sequences mclude, but are not limited to, the GCG program package (Devereux, J , et al , Nucleic Acids Research 12(1) 387 (1984)), BLASTP, BLASTN, and FASTA (Atschul, S F et al , J Molec Biol 215 403-410 (1990) The BLAST X program is publicly available from NCBI and other sources (BLAST Manual, Altschul, S , et al , NCBI NLM NM Bethesda, MD 20894, Altschul, S , et al , J Mol Biol 215 403-410 (1990) The well known Smith Waterman algonthm may also be used to determine identity

Preferred parameters for polypeptide sequence companson include the following 1) Algonthm Needleman and Wunsch, J Mol Biol 48 443-453 (1970) Companson matrix BLOSSUM62 from Hentikoff and Hentikoff, Proc Natl Acad Sci USA 89 10915-10919 (1992) Gap Penalty 12 Gap Length Penalty 4

A program useful with these parameters is publicly available as the "gap" program from Genetics Computer Group, Madison WI The aforementioned parameters are the default

4 parameters for polypeptide comparisons (along with no penalty for end gaps)

Preferred parameters for polynucleotide comparison include the following 1) Algonthm Needleman and Wunsch, J Mol Biol 48 443-453 (1970) Companson matnx matches = +10, mismatch = 0

Gap Penalty 50 Gap Length Penalty 3

A program useful with these parameters is publicly available as the "gap" program from Genetics Computer Group, Madison WI The aforementioned parameters are the default parameters for polynucleotide compansons

Preferred polynucleotide embodiments further mclude an isolated polynucleotide comprising a polynucleotide havmg at least a 50,60, 70, 80, 85, 90, 95, 97 or 100% identity to a polynucleotide reference sequence of SEQ ID NO 1, wherem said reference sequence may be identical to the sequence of SEQ ID NO 1 or may mclude up to a certain integer number of nucleotide alterations as compared to the reference sequence, wherein said alterations are selected from the group consisting of at least one nucleotide deletion, substitution, including transition and transversion, or insertion, and wherein said alterations may occur at the 5' or 3' terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, mterspersed either individually among the nucleotides m the reference sequence or m one or more contiguous groups within the reference sequence, and wherem said number of nucleotide alterations is determined by multiplying the total number of nucleotides in SEQ ID NO 1 by the numencal percent of the respective percent identity and subtractmg that product from said total number of nucleotides in SEQ ID NO 1, or

nn £ xn - (xn ^• y),

wherein nn is the number of nucleotide alterations, xn is the total number of nucleotides in SEQ ID NO 1, and y is 0 50 for 50%, 0 60 for 60%, 0 70 for 70%, 0 80 for 80%, 0 85 for 85%, 0 90 for 90%, 0 95 for 95%, 0 97 for 97% or 1 00 for 100%, and wherem any non-integer product of xn and y is rounded down to the nearest integer prior to subtracting it from xn Alterations of a polynucleotide sequence encoding the polypeptide of SEQ ID NO 2 may create nonsense, missense or frameshift mutations in this coding sequence and thereby alter the polypeptide encoded by the polynucleotide following such alterations

5 Preferred polypeptide embodiments further include an isolated polypeptide comprising a polypeptide having at least a 50,60, 70, 80, 85, 90, 95, 97 or 100% identity to a polypeptide reference sequence of SEQ ID NO:2, wherein said reference sequence may be identical to the sequence of SEQ ID NO: 2 or may include up to a certain integer number of amino acid alterations as compared to the reference sequence, wherein said alterations are selected from the group consisting of at least one amino acid deletion, substitution, including conservative and non- conservative substitution, or insertion, and wherein said alterations may occur at the amino- or carboxy-terminal positions of the reference polypeptide sequence or anywhere between those terminal positions, interspersed either individually among the amino acids in the reference sequence or in one or more contiguous groups within the reference sequence, and wherein said number of amino acid alterations is determined by multiplying the total number of amino acids in SEQ ID NO:2 by the numerical percent of the respective percent identity and subtracting that product from said total number of amino acids in SEQ ID NO:2, or:

na £ xa - (xa ^• y),

wherein na is the number of amino acid alterations, xa is the total number of amino acids in SEQ ID NOJ, and y is 0.50 for 50%, 0.60 for 60%, 0.70 for 70%, 0.80 for 80%, 0.85 for 85%, 0.90 for 90%, 0.95 for 95%, 0.97 for 97% or 1.00 for 100%, and wherein any non-integer product of xa and y is rounded down to the nearest integer prior to subtracting it from xa.

Polypeptides of the Invention In one aspect, the present invention relates to CBDAME07 polypeptides (or CBDAME07 proteins). The CBDAME07 polypeptides include the polypeptide of SEQ ID NOJ; as well as polypeptides comprising the amino acid sequence of SEQ ID NO: 2; and polypeptides comprising the amino acid sequence which have at least 80% identity to that of SEQ ID NOJ over its entire length, and still more preferably at least 90% identity, and even still more preferably at least 95% identity to SEQ ID NO: 2. Furthermore, those with at least 97-99% are highly preferred. Also included within CBDAME07 polypeptides are polypeptides having the amino acid sequence which have at least 80% identity to the polypeptide having the amino acid sequence of SEQ ID NOJ over its entire length, and still more preferably at least 90% identity, and still more preferably at least 95% identity to SEQ ID NO 2 Furthermore, those with at least 97-99% are highly preferred Preferably CBDAME07 polypeptide exhibit at least one biological activity of CBDAME07

The CBDAME07 polypeptides may be m the form of the "mature" protem or may be a part of a larger protem such as a fusion protem It is often advantageous to mclude an additional ammo acid sequence which contams secretory or leader sequences, pro-sequences, sequences which aid in purification such as multiple histidine residues, or an additional sequence for stability during recombinant production

Fragments of the CBDAME07 polypeptides are also mcluded in the invention A fragment is a polypeptide having an ammo acid sequence that entirely is the same as part, but not all, of the amino acid sequence of the aforementioned CBDAME07 polypeptides As with CBDAME07 polypeptides, fragments may be "free-standmg," or compnsed within a larger polypeptide of which they form a part or region, most preferably as a single continuous region Representative examples of polypeptide fragments of the invention, include, for example, fragments from about ammo acid number 1-20, 21-40, 41-60, 61- 80, 81-100, and 101 to the end of CBDAME07 polypeptide In this context "about" mcludes the particularly recited ranges larger or smaller by several, 5, 4, 3, 2 or 1 ammo acid at either extreme or at both extremes

Preferred fragments mclude, for example, truncation polypeptides havmg the ammo acid sequence of CBDAME07 polypeptides, except for deletion of a continuous senes of residues that mcludes the amino terminus, or a continuous senes of residues that mcludes the carboxyl terminus or deletion of two continuous senes of residues, one mcludmg the ammo terminus and one mcludmg the carboxyl terminus Also preferred are fragments characterized by structural or functional attributes such as fragments that compnse alpha-helix and alpha-helix forming regions, beta-sheet and beta-sheet- forming regions, turn and turn-forming regions, coil and coil-forming regions, hydrophihc regions, hydrophobic regions, alpha amphipathic regions, beta amphipathic regions, flexible regions, surface- forming regions, substrate binding region, and high antigemc mdex regions Other preferred fragments are biologically active fragments Biologically active fragments are those that mediate CBDAME07 activity, including those with a similar activity or an improved activity, or with a decreased undesirable activity Also included are those that are antigemc or lmmunogenic m an animal, especially in a human Preferably, all of these polypeptide fragments retain the biological activity of the CBDAME07 , mcludmg antigemc activity Vanants of the defined sequence and fragments also form part of the present invention Preferred vanants are those that vary from the referents by conservative ammo acid substitutions — l e , those that substitute a residue with another of like charactenstics Typical such substitutions are among Ala, Val, Leu and lie, among Ser and Thr, among the acidic residues Asp and Glu, among Asn and Gin, and among the basic residues Lys and Arg, or aromatic residues Phe and Tyr

7 Particularly preferred are variants in which several, 5-10, 1-5, or 1-2 amino acids are substituted, deleted, or added in any combination.

The CBDAME07 polypeptides of the invention can be prepared in any suitable manner. Such polypeptides include isolated naturally occurring polypeptides, recombinantly produced polypeptides, synthetically produced polypeptides, or polypeptides produced by a combination of these methods. Means for preparing such polypeptides are well understood in the art.

Polynucleotides of the Invention

Another aspect of the invention relates to CBDAME07 polynucleotides. CBDAME07 polynucleotides include isolated polynucleotides which encode the CBDAME07 polypeptides and fragments, and polynucleotides closely related thereto. More specifically, CBDAME07 polynucleotide of the invention include a polynucleotide comprising the nucleotide sequence contained in SEQ ID NO: 1 encoding a CBDAME07 polypeptide of SEQ ID NO: 2, and polynucleotide having the particular sequence of SEQ ID NO: 1. CBDAME07 polynucleotides further include a polynucleotide comprising a nucleotide sequence that has at least 80% identity over its entire length to a nucleotide sequence encoding the CBDAME07 polypeptide of SEQ ID NOJ, and a polynucleotide comprising a nucleotide sequence that is at least 80% identical to of SEQ ID NO: 1 over its entire length. In this regard, polynucleotides at least 90% identical are particularly preferred, and those with at least 95% are especially preferred. Furthermore, those with at least 97% are highly preferred and those with at least 98-99% are most highly preferred, with at least 99% being the most preferred. Also included under CBDAME07 polynucleotides are a nucleotide sequence which has sufficient identity to a nucleotide sequence contained in SEQ ID NO:l to hybridize under conditions useable for amplification or for use as a probe or marker. The invention also provides polynucleotides which are complementary to such CBDAME07 polynucleotides. The CBDAME07 of the invention is structurally related to other proteins of the protein transporter sec61 family, as shown by the results of sequencing the cDNA of Table 1 (SEQ ID NO: 1) encoding human CBDAME07. The cDNA sequence of SEQ ID NO: 1 contains an open reading frame (nucleotide number 91 to 294) encoding a polypeptide of 68 amino acids of SEQ ID NOJ. The amino acid sequence of Table 2 (SEQ ID NOJ) has about 99% identity (using FASTA) in 68 amino acid residues with canine and murine Sec61 gamma subunit (E. Hartmann, et al. Nature. 1994, 367: 654- 657). The nucleotide sequence of Table 1 (SEQ ID NO: 1) has about 70.1% identity (using FASTA) in 482 nucleotide residues with canine and murine Sec61 gamma subunit (E. Hartmann, et al. Nature 1994, 367: 654-657). Thus, CBDAME07 polypeptides and polynucleotides of the present invention are expected to have, inter aha, similar biological functions/properties to their homologous polypeptides and polynucleotides, and their utility is obvious to anyone skilled in the art.

Table 1'

1 TCGACCCGGA TTCCGGTTCC GGTGGGCTCC ATCAGCAAGC TCCAGTGCTA

51 CGTGTCCCTG GCATTTTAGG TGTCGGTTGG GTAGGCAGTC ATGGATCAGG

101 TAATGCAGTT TGTTGAGCCA AGTCGGCAGT TTGTAAAGGA CTCCATTCGG

151 CTGGTTAAAA GATGCACTAA ACCTGATAGA AAAGAATTCC AGAAGATTGC

201 CATGGCAACA GCAATAGGAT TTGCTATAAT GGGATTCATT GGCTTCTTTG

251 TGAAATTGAT CCATATTCCT ATTAATAACA TCATTGTTGG TGGCTGAATA

301 CATTTTGGAA GAGAGTTTTT CATCTTAGAG ATTGGTGAAC AAGTGTGAGG

351 GTGTGAGAAA CTCACAGAAT ACAAATTTGC CTGTATGTTT TGTGGGTTTT

401 TTTTTTTCCT TTCAAGATGT TTTCTATTTC TAAATTAAAG TAATTTCAAA

451 ATAAAAAAAA AAAAAAAAAA AAAAAAAAAA AA

A nucleotide sequence of a human CBDAME07 (SEQ ID NO: 1).

Table 2^b

1 MDQVMQFVEP SRQFVKDSIR LVKRCTKPDR KEFQKIAMAT AIGFAIMGFI

51 GFFVKLIHIP I NIIVGG

An amino acid sequence of a human CBDAME07 (SEQ ID NO: 2).

One polynucleotide of the present invention encoding CBDAME07 may be obtained using standard cloning and screening, from a cDNA library derived from mRNA in cells of human cord blood using the expressed sequence tag (EST) analysis (Adams, M.D., et al. Science (1991) 252: 1651- 1656; Adams, M.D. et al, Nature, (1992) 55:632-634; Adams, M.D., et al, Nature (1995) 377 Supp:3-174). Polynucleotides of the invention can also be obtained from natural sources such as genomic DNA libraries or can be synthesized usmg well known and commercially available techniques

The nucleotide sequence encodmg CBDAME07 polypeptide of SEQ ID NO 2 may be identical to the polypeptide encodmg sequence contained m Table 1 (nucleotide number 91 to 294 of SEQ ID NO 1), or it may be a sequence, which as a result of the redundancy (degeneracy) of the genetic code, also encodes the polypeptide of SEQ ID NO 2

When the polynucleotides of the invention are used for the recombinant production of CBDAME07 polypeptide, the polynucleotide may mclude the coding sequence for the mature polypeptide or a fragment thereof, by itself, the coding sequence for the mature polypeptide or fragment m readmg frame with other coding sequences, such as those encoding a leader or secretory sequence, a pre-, or pro- or prepro- protem sequence, or other fusion peptide portions For example, a marker sequence which facilitates punfication of the fused polypeptide can be encoded In certain preferred embodiments of this aspect of the invention, the marker sequence is a hexa-histidine peptide, as provided m the pQE vector (Qiagen, Inc ) and descnbed in Gentz et al , Proc NatlAcad Set USA (1989) 86 821- 824, or is an HA tag The polynucleotide may also contain non-coding 5 ' and 3 ' sequences, such as transcnbed, non-translated sequences, splicing and polyadenylation signals, nbosome binding sites and sequences that stabilize mRNA

Further preferred embodiments are polynucleotides encodmg CBDAME07 vanants compnse the ammo acid sequence CBDAME07 polypeptide of Table 2 (SEQ ID NO 2) m which several, 5-10, 1-5, 1-3, 1-2 or 1 ammo acid residues are substituted, deleted or added, m any combination

The present invention further relates to polynucleotides that hybndize to the herem above- descnbed sequences In this regard, the present invention especially relates to polynucleotides which hybndize under stringent conditions to the herem above-descnbed polynucleotides As herein used, the term "stringent conditions" means hybndization will occur only if there is at least 80%, and preferably at least 90%, and more preferably at least 95%, yet even more preferably 97-99% identity between the sequences

Polynucleotides of the invention, which are identical or sufficiently identical to a nucleotide sequence contained m SEQ ID NO 1 or a fragment thereof, may be used as hybndization probes for cDNA and genomic DNA, to isolate full-length cDNAs and genomic clones encodmg CBDAME07 polypeptide and to isolate cDNA and genomic clones of other genes (mcludmg genes encodmg homologs and orthologs from species other than human) that have a high sequence similanty to the CBDAME07 gene Such hybndization techniques are known to those of skill in the art Typically these nucleotide sequences are 80% identical, preferably 90% identical, more preferably 95% identical to that of the referent The probes generally will compnse at least 15 nucleotides Preferably, such probes will have at

10 least 30 nucleotides and may have at least 50 nucleotides. Particularly preferred probes will range between 30 and 50 nucleotides.

In one embodiment, to obtain a polynucleotide encoding CBDAME07 polypeptide, including homologs and orthologs from species other than human, comprises the steps of screening an appropriate library under stingent hybridization conditions with a labeled probe having the SEQ ID NO: 1 or a fragment thereof; and isolating full-length cDNA and genomic clones containing said polynucleotide sequence. Thus in another aspect, CBDAME07 polynucleotides of the present invention further include a nucleotide sequence comprising a nucleotide sequence that hybridize under stringent condition to a nucleotide sequence having SEQ ID NO: 1 or a fragment thereof. Also included with CBDAME07 polypeptides are polypeptide comprising amino acid sequence encoded by nucleotide sequence obtained by the above hybridization condition. Such hybridization techniques are well known to those of skill in the art. Stringent hybridization conditions are as defined above or, alternatively, conditions under overnight incubation at 42°C in a solution comprising: 50% formamide, 5xSSC (150mM NaCl, 15mM trisodium citrate), 50 mM sodium phosphate (pH7.6), 5x Denhardt's solution, 10 % dextran sulfate, and 20 microgram ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0. Ix SSC at about 65°C.

The polynucleotides and polypeptides of the present invention may be employed as research reagents and materials for discovery of treatments and diagnostics to animal and human disease.

Vectors, Host Cells, Expression

The present invention also relates to vectors which comprise a polynucleotide or polynucleotides of the present invention, and host cells which are genetically engineered with vectors of the invention and to the production of polypeptides of the invention by recombinant techniques. Cell-free translation systems can also be employed to produce such proteins using RNAs derived from the DNA constructs of the present invention.

For recombinant production, host cells can be genetically engineered to incorporate expression systems or portions thereof for polynucleotides of the present invention. Introduction of polynucleotides into host cells can be effected by methods described in many standard laboratory manuals, such as Davis et a\., BASIC METHODS IN MOLECULAR BIOLOGY (1986) and Sambrook et al., MOLECULAR CLONING: A LABORATORY MANUAL, 2nd Ed, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989) such as calcium phosphate transfection, DEAE-dextran mediated transfection, transvection, microinjection, cationic lipid-mediated transfection, electroporation, transduction, scrape loading, ballistic introduction or infection.

11 Representative examples of appropnate hosts mclude bactenal cells, such as streptococci, staphylococci, E coh, Streptomyces and Bacillus subtihs cells, fungal cells, such as yeast cells and Aspergillus cells, insect cells such as Drosophila S2 and Spodoptera Sf9 cells, animal cells such as CHO, COS, HeLa, C127, 3T3, BHK, HEK 293 and Bowes melanoma cells, and plant cells A great vanety of expression systems can be used Such systems mclude, among others, chromosomal, episomal and virus-denved systems, e g , vectors denved from bactenal plasπuds, from bactenophage, from transposons, from yeast episomes, from insertion elements, from yeast chromosomal elements, from viruses such as baculoviruses, papova viruses, such as SV40, vaccinia viruses, adenoviruses, fowl pox viruses, pseudorabies viruses and retrovinises, and vectors denved from combinations thereof, such as those denved from plasmid and bactenophage genetic elements, such as cosmids and phagemids The expression systems may contain control regions that regulate as well as engender expression Generally, any system or vector suitable to maintain, propagate or express polynucleotides to produce a polypeptide m a host may be used The appropnate nucleotide sequence may be inserted into an expression system by any of a vanety of well-known and routine techmques, such as, for example, those set forth m Sambrook et al , MOLECULAR CLONING, A LABORATORY MANUAL (supra)

For secretion of the translated protem into the lumen of the endoplasmic reticulum, into the penplasmic space or into the extracellular environment, appropnate secretion signals may be incorporated into the desired polypeptide These signals may be endogenous to the polypeptide or they may be heterologous signals

If the CBDAME07 polypeptide is to be expressed for use in screemng assays, generally, it is preferred that the polypeptide be produced at the surface of the cell In this event, the cells may be harvested prior to use m the screening assay If CBDAME07 polypeptide is secreted mto the medium, the medium can be recovered m order to recover and puπfy the polypeptide, if produced intracellularly, the cells must first be lysed before the polypeptide is recovered

CBDAME07 polypeptides can be recovered and purified from recombinant cell cultures by well-known methods mcludmg ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography Most preferably, high performance liquid chromatography is employed for punfication Well known techmques for refoldmg proteins may be employed to regenerate active conformation when the polypeptide is denatured during isolation and or punfication

12 Diagnostic Assays

This invention also relates to the use of CBDAME07 polynucleotides for use as diagnostic reagents Detection of a mutated form of CBDAME07 gene associated with a dysfunction will provide a diagnostic tool that can add to or define a diagnosis of a disease or susceptibility to a disease which results from under-expression, over-expression or altered expression of CBDAME07 Individuals carrying mutations m the CBDAME07 gene may be detected at the DNA level by a vanety of techmques

Nucleic acids for diagnosis may be obtained from a subject's cells, such as from blood, urine, saliva, tissue biopsy or autopsy matenal The genomic DNA may be used directly for detection or may be amplified enzymatically by usmg PCR or other amplification techmques pnor to analysis RNA or cDNA may also be used in similar fashion Deletions and insertions can be detected by a change m size of the amplified product in companson to the normal genotype Pomt mutations can be identified by hybndizing amplified DNA to labeled CBDAME07 nucleotide sequences Perfectly matched sequences can be distinguished from mismatched duplexes by RNase digestion or by differences m melting temperatures DNA sequence differences may also be detected by alterations m electiophoretic mobility of DNA fragments m gels, with or without denaturing agents, or by direct DNA sequencmg See, e g , Myers et al , Science (1985) 230 1242 Sequence changes at specific locations may also be revealed by nuclease protection assays, such as RNase and S 1 protection or the chemical cleavage method See Cotton et al , Proc Natl Acad Sci USA ( 1985) 85 4397-4401 In another embodiment, an array of ohgonucleotides probes compnsing CBDAME07 nucleotide sequence or fragments thereof can be constructed to conduct efficient screemng of e g , genetic mutations Array technology methods are well known and have general applicability and can be used to address a vanety of questions m molecular genetics mcludmg gene expression, genetic linkage, and genetic vanability (See for example M Chee et al , Science, Vol 274, pp 610-613 (1996)) The diagnostic assays offer a process for diagnosing or determining a susceptibility to cancer,

AIDS, metabolic disorders, and IDDM through detection of mutation m the CBDAME07 gene by the methods desenbed

In addition, cancer, AIDS, metabolic disorders, and IDDM , can be diagnosed by methods comprising determining from a sample denved from a subject an abnormally decreased or increased level of CBDAME07 polypeptide or CBDAME07 mRNA Decreased or increased expression can be measured at the RNA level using any of the methods well known m the art for the quantitation of polynucleotides, such as, for example, PCR, RT-PCR, RNase protection, Northern blotting and other hybndization methods Assay techmques that can be used to determme levels of a protem, such as an CBDAME07 polypeptide, m a sample denved from a host are well-

13 known to those of skill m the art Such assay methods mclude radioimmunoassays, competitive-binding assays, Western Blot analysis and ELISA assays

Thus in another aspect, the present mvention relates to a diagonostic kit for a disease or suspectabihty to a disease, particularly cancer, AIDS, metabolic disorders, and IDDM , which compnses

(a) a CBDAME07 polynucleotide, preferably the nucleotide sequence of SEQ ID NO 1, or a fragment thereof ,

(b) a nucleotide sequence complementary to that of (a),

(c) a CBDAME07 polypeptide, preferably the polypeptide of SEQ ID NO 2, or a fragment thereof, or

(d) an antibody to a CBDAME07 polypeptide, preferably to the polypeptide of SEQ ID NO 2 It will be appreciated that in any such kit, (a), (b), (c) or (d) may comprise a substantial component

Chromosome Assays

The nucleotide sequences of the present mvention are also valuable for chromosome identification The sequence is specifically targeted to and can hybndize with a particular location on an individual human chromosome The mapping of relevant sequences to chromosomes accordmg to the present mvention is an important first step m correlating those sequences with gene associated disease Once a sequence has been mapped to a precise chromosomal location, the physical position of the sequence on the chromosome can be correlated with genetic map data Such data are found, for example, m V McKusick, Mendehan Inhentance m Man (available on lme through Johns Hopkins Umversity Welch Medical Library) The relationship between genes and diseases that have been mapped to the same chromosomal region are then identified through linkage analysis (comhentance of physically adjacent genes)

The differences in the cDNA or genomic sequence between affected and unaffected individuals can also be determined If a mutation is observed m some or all of the affected individuals but not m any normal individuals, then the mutation is likely to be the causative agent of the disease

Antibodies

The polypeptides of the mvention or their fragments or analogs thereof, or cells expressing them can also be used as lmmunogens to produce antibodies lmmunospecific for the CBDAME07

14 polypeptides The term "lrnmunospecrfic" means that the antibodies have substantiall greater affinity for the polypeptides of the mvention than then affinity for other related polypeptides m the pnor art

Antibodies generated against the CBDAME07 polypeptides can be obtained by administering the polypeptides or epitope-beaπng fragments, analogs or cells to an animal, preferably a nonhuman, usmg routine protocols For preparation of monoclonal antibodies, any technique which provides antibodies produced by continuous cell line cultures can be used Examples mclude the hybndoma technique (Kohler, G and Milstein, C , Nature (1975) 256 495-497), the tnoma technique, the human B-cell hybndoma technique (Kozbor et al , Immunology Today (1983) 472) and the EBV-hybndoma technique (Cole et al , MONOCLONAL ANTIBODIES AND CANCER THERAPY, pp 77-96, Alan R Liss, Inc , 1985)

Techmques for the production of smgle chain antibodies (U S Patent No 4,946,778) can also be adapted to produce smgle chain antibodies to polypeptides of this mvention Also, transgenic mice, or other organisms mcludmg other mammals, may be used to express humanized antibodies

The above-descnbed antibodies may be employed to isolate or to identify clones expressmg the polypeptide or to punfy the polypeptides by affinity chromatography

Antibodies against CBDAME07 polypeptides may also be employed to treat cancer, AIDS, metabolic disorders, and IDDM , among others

Vaccines Another aspect of the mvention relates to a method for mducmg an lmmunological response in a mammal which compnses moculatmg the mammal with CBDAME07 polypeptide, or a fragment thereof, adequate to produce antibody and or T cell immune response to protect said animal from cancer, AIDS, metabolic disorders, and IDDM , among others Yet another aspect of the invention relates to a method of mducmg lmmunological response m a mammal which comprises, dehvermg CBDAME07 polypeptide via a vector directmg expression of CBDAME07 polynucleotide in vivo m order to mduce such an lmmunological response to produce antibody to protect said animal from diseases

Further aspect of the mvention relates to an immunological/vaccine formulation (composition) which, when introduced into a mammalian host, induces an lmmunological response in that mammal to a CBDAME07 polypeptide wherem the composition comprises a CBDAME07 polypeptide or CBDAME07 gene The vaccine formulation may further compnse a suitable carrier Since CBDAME07 polypeptide may be broken down in the stomach, it is preferably administered parenterally (including subcutaneous, intramuscular, intravenous, mtradermal etc injection) Formulations suitable for parenteral administration mclude aqueous and non-aqueous

15 sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation instonic with the blood of the recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents or thickening agents. The formulations may be presented in unit-dose or multi-dose containers, for example, sealed ampoules and vials and may be stored in a freeze-dried condition requiring only the addition of the sterile liquid carrier immediately prior to use. The vaccine formulation may also include adjuvant systems for enhancing the immunogenicity of the formulation, such as oil-in water systems and other systems known in the art. The dosage will depend on the specific activity of the vaccine and can be readily determined by routine experimentation.

Screening Assays

The CBDAME07 polypeptide of the present invention may be employed in a screening process for compounds which activate (agonists) or inhibit activation of (antagonists, or otherwise called inhibitors) the CBDAME07 polypeptide of the present invention. Thus, polypeptides of the invention may also be used to assess identify agonist or antagonists from, for example, cells, cell-free preparations, chemical libraries, and natural product mixtures. These agonists or antagonists may be natural or modified substrates, ligands, receptors, enzymes, etc, as the case may be, of the polypeptide of the present invention; or may be structural or functional mimetics of the polypeptide of the present invention. See Coligan et al. , Current Protocols in Immunology l(2):Chapter 5 (1991). CBDAME07 polypeptides are responsible for many biological functions, including many pathologies. Accordingly, it is desirous to find compounds and drugs which stimulate CBDAME07 polypeptide on the one hand and which can inhibit the function of CBDAME07 polypeptide on the other hand. In general, agonists are employed for therapeutic and prophylactic purposes for such conditions as cancer, AIDS, metabolic disorders, and IDDM . Antagonists may be employed for a variety of therapeutic and prophylactic purposes for such conditions as cancer, AIDS, metabolic disorders, and IDDM .

In general, such screening procedures may involve using appropriate cells which express the CBDAME07 polypeptide or respond to CBDAME07 polypeptide of the present invention. Such cells include cells from mammals, yeast, Drosophila or E. coli. Cells which express the CBDAME07 polypeptide (or cell membrane containing the expressed polypeptide) or respond to CBDAME07 polypeptide are then contacted with a test compound to observe binding, or stimulation or inhibition of a functional response. The ability of the cells which were contacted with the candidate compounds is compared with the same cells which were not contacted for CBDAME07 activity.

16 The assays may simply test binding of a candidate compound wherein adherence to the cells bearing the CBDAME07 polypeptide is detected by means of a label directly or indirectly associated with the candidate compound or m an assay involving competition with a labeled competitor Further, these assays may test whether the candidate compound results in a signal generated by activation of the CBDAME07 polypeptide, using detection systems appropnate to the cells beanng the CBDAME07 polypeptide Inhibitors of activation are generally assayed m the presence of a known agomst and the effect on activation by the agomst by the presence of the candidate compound is observed

Further, the assays may simply compnse the steps of mixing a candidate compound with a solution contammg a CBDAME07 polypeptide to form a mixture, measurmg CBDAME07 activity m the mixture, and comparing the CBDAME07 activity of the mixture to a standard

The CBDAME07 cDNA, protein and antibodies to the protem may also be used to configure assays for detectmg the effect of added compounds on the production of CBDAME07 mRNA and protein m cells For example, an ELISA may be constructed for measunng secreted or cell associated levels of CBDAME07 protem using monoclonal and polyclonal antibodies by standard methods known in the art, and this can be used to discover agents which may inhibit or enhance the production of CBDAME07 (also called antagonist or agomst, respectively) from suitably manipulated cells or tissues

The CBDAME07 protein may be used to identify membrane bound or soluble receptors, if any, through standard receptor bindmg techmques known in the art These mclude, but are not limited to, hgand bindmg and crosshnking assays in which the CBDAME07 is labeled with a radioactive isotope (eg 1251), chemically modified (eg biotinylated), or fused to a peptide sequence suitable for detection or punfication, and incubated with a source of the putative receptor (cells, cell membranes, cell supernatants, tissue extracts, bodily fluids) Other methods mclude biophysical techmques such as surface plasmon resonance and spectroscopy In addition to being used for purification and cloning of the receptor, these binding assays can be used to identify agonists and antagonists of CBDAME07 which compete with the binding of CBDAME07 to its receptors, if any Standard methods for conducting screemng assays are well understood in the art Examples of potential CBDAME07 polypeptide antagonists mclude antibodies or, m some cases, oligonucleotides or proteins which are closely related to the gands, substrates, receptors, enzymes, etc , as the case may be, of the CBDAME07 polypeptide, e g , a fragment of the gands, substrates, receptors, enzymes, etc , or small molecules which bmd to the polypetide of the present mvention but do not elicit a response, so that the activity of the polypeptide is prevented

17 Thus in another aspect, the present invention relates to a screening kit for identifying agonists, antagonists, Hgands, receptors, substrates, enzymes, etc. for CBDAME07 polypeptides; or compounds which decrease or enhance the production of CBDAME07 polypeptides, which comprises: (a) a CBDAME07 polypeptide, preferably that of SEQ ID NOJ;

(b) a recombinant cell expressing a CBDAME07 polypeptide, preferably that of SEQ ID NOJ;

(c) a cell membrane expressing a CBDAME07 polypeptide; preferably that of SEQ ID NO: 2; or

(d) antibody to a CBDAME07 polypeptide, preferably that of SEQ ID NO: 2.

It will be appreciated that in any such kit, (a), (b), (c) or (d) may comprise a substantial component.

Prophylactic and Therapeutic Methods

This invention provides methods of treating abnormal conditions such as, cancer, AIDS, metabolic disorders, and IDDM , related to both an excess of and insufficient amounts of CBDAME07 polypeptide activity.

If the activity of CBDAME07 polypeptide is in excess, several approaches are available. One approach comprises administering to a subject an inhibitor compound (antagonist) as hereinabove described along with a pharmaceutically acceptable carrier in an amount effective to inhibit the function of the CBDAME07 polypeptide, such as, for example, by blocking the binding of hgands, substrates, receptors, enzymes, etc, or by inhibiting a second signal, and thereby alleviating the abnormal condition. In another approach, soluble forms of CBDAME07 polypeptides still capable of binding the ligand, substrate, enzymes, receptors, etc. in competition with endogenous CBDAME07 polypeptide may be administered. Typical embodiments of such competitors comprise fragments of the CBDAME07 polypeptide. In still another approach, expression of the gene encoding endogenous CBDAME07 polypeptide can be inhibited using expression blocking techniques. Known such techniques involve the use of antisense sequences, either internally generated or separately administered. See, for example, O'Connor, J Neurochem (1991) 56:560 in Oligodeoxynucleotides as Antisense Inhibitors of Gene Expression, CRC Press, Boca Raton, FL (1988). Alternatively, oligonucleotides which form triple helices with the gene can be supplied. See, for example, Lee et al, Nucleic Acids Res (1979) 6:3073; Cooney et al, Science (1988) 241:456; Dervan ef α/, Science (1991) 251: 1360. These oligomers can be administered per se or the relevant oligomers can be expressed in vivo.

For treating abnormal conditions related to an under-expression of CBDAME07 and its activity, several approaches are also available. One approach comprises administering to a subject a

18 therapeutically effective amount of a compound which activates CBDAME07 polypeptide, i.e, an agonist as described above, in combination with a pharmaceutically acceptable carrier, to thereby alleviate the abnormal condition. Alternatively, gene therapy may be employed to effect the endogenous production of CBDAME07 by the relevant cells in the subject. For example, a polynucleotide of the invention may be engineered for expression in a replication defective retroviral vector, as discussed above. The retroviral expression construct may then be isolated and introduced into a packaging cell transduced with a retroviral plasmid vector containing RNA encoding a polypeptide of the present invention such that the packaging cell now produces infectious viral particles containing the gene of interest. These producer cells may be administered to a subject for engineering cells in vivo and expression of the polypeptide in vivo. For overview of gene therapy, see Chapter 20, Gene Therapy and other Molecular Genetic-based Therapeutic Approaches, (and references cited therein) in Human Molecular Genetics, T Strachan and A P Read, BIOS Scientific Publishers Ltd (1996). Another approach is to administer a therapeutic amount of CBDAME07 polypeptides in combination with a suitable pharmaceutical carrier.

Formulation and Administration

Peptides, such as the soluble form of CBDAME07 polypeptides, and agonists and antagonist peptides or small molecules, may be formulated in combination with a suitable pharmaceutical carrier. Such formulations comprise a therapeutically effective amount of the polypeptide or compound, and a pharmaceutically acceptable carrier or excipient. Such carriers include but are not limited to, saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations thereof. Formulation should suit the mode of administration, and is well within the skill of the art. The invention further relates to pharmaceutical packs and kits comprising one or more containers filled with one or more of the ingredients of the aforementioned compositions of the invention. Polypeptides and other compounds of the present invention may be employed alone or in conjunction with other compounds, such as therapeutic compounds.

Preferred forms of systemic administration of the pharmaceutical compositions include injection, typically by intravenous injection. Other injection routes, such as subcutaneous, intramuscular, or intraperitoneal, can be used. Alternative means for systemic administration include transmucosal and transdermal administration using penetrants such as bile salts or fusidic acids or other detergents. In addition, if properly formulated in enteric or encapsulated formulations, oral administration may also be possible. Administration of these compounds may also be topical and/or localized, in the form of salves, pastes, gels and the like.

19 The dosage range required depends on the choice of peptide, the route of administration, the nature of the formulation, the nature of the subject's condition, and the judgment of the attending practitioner. Suitable dosages, however, are in the range of 0 J - 100 μg/kg of subject. Wide variations in the needed dosage, however, are to be expected in view of the variety of compounds available and the differing efficiencies of various routes of administration. For example, oral administration would be expected to require higher dosages than administration by intravenous injection. Variations in these dosage levels can be adjusted using standard empirical routines for optimization, as is well understood in the art.

Polypeptides used in treatment can also be generated endogenously in the subject, in treatment modalities often referred to as "gene therapy" as described above. Thus, for example, cells from a subject may be engineered with a polynucleotide, such as a DNA or RNA, to encode a polypeptide ex vivo, and for example, by the use of a retroviral plasmid vector. The cells are then introduced into the subject.

All publications, including but not limited to patents and patent applications, cited in this specification are herein incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference herein as though fully set forth.

20 SEQUENCE LISTING

(1) GENERAL INFORMATION

(i) APPLICANT: WU, JI-SHENG FU, GANG MAO, MAO

SHEN, YU

(ii) TITLE OF THE INVENTION: Human Secβl Gamma Subunit Gene: CBDAME07

(iii) NUMBER OF SEQUENCES: 2

(iv) CORRESPONDENCE ADDRESS:

(A) ADDRESSEE: RATNER & PRESTIA (B) STREET: P.O. BOX 980

(C) CITY: VALLEY FORGE

(D) STATE: PA

(E) COUNTRY: USA

(F) ZIP: 19482

(v) COMPUTER READABLE FORM:

(A) MEDIUM TYPE: Diskette

(B) COMPUTER: IBM Compatible

(C) OPERATING SYSTEM: DOS (D) SOFTWARE: FastSEQ for Windows Version 2.0

(vi) CURRENT APPLICATION DATA:

(A) APPLICATION NUMBER: TO BE ASSIGND

(B) FILING DATE: (C) CLASSIFICATION: UNKNOWN

(vii) PRIOR APPLICATION DATA:

(A) APPLICATION NUMBER:

(B) FILING DATE:

(viii) ATTORNEY/AGENT INFORMATION:

21 (A) NAME: PRESTIA, PAUL F

(B) REGISTRATION NUMBER: 23,031

(C) REFERENCE/DOCKET NUMBER: GP-70411

(ix) TELECOMMUNICATION INFORMATION:

(A) TELEPHONE: 610-407-0700

(B) TELEFAX: 610-407-0701

(C) TELEX: 846169

(2) INFORMATION FOR SEQ ID NO : 1 :

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 482 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

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

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

TCGACCCGGA TTCCGGTTCC GGTGGGCTCC ATCAGCAAGC TCCAGTGCTA CGTGTCCCTG 60

GCATTTTAGG TGTCGGTTGG GTAGGCAGTC ATGGATCAGG TAATGCAGTT TGTTGAGCCA 120

AGTCGGCAGT TTGTAAAGGA CTCCATTCGG CTGGTTAAAA GATGCACTAA ACCTGATAGA 180 AAAGAATTCC AGAAGATTGC CATGGCAACA GCAATAGGAT TTGCTATAAT GGGATTCATT 240

GGCTTCTTTG TGAAATTGAT CCATATTCCT ATTAATAACA TCATTGTTGG TGGCTGAATA 300

CATTTTGGAA GAGAGTTTTT CATCTTAGAG ATTGGTGAAC AAGTGTGAGG GTGTGAGAAA 360

CTCACAGAAT ACAAATTTGC CTGTATGTTT TGTGGGTTTT TTTTTTTCCT TTCAAGATGT 420

TTTCTATTTC TAAATTAAAG TAATTTCAAA ATAAAAAAAA AAAAAAAAAA AAAAAAAAAA 480 AA 482

(2) INFORMATION FOR SEQ ID NO: 2:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 68 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

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

Met Asp Gin Val Met Gin Phe Val Glu Pro Ser Arg Gin Phe Val Lys

22 1 5 10 15 sp Ser lie Arg Leu Val Lys Arg Cys Thr Lys Pro Asp Arg Lys Glu

20 25 30

Phe Gin Lys lie Ala Met Ala Thr Ala l ie Gly Phe Ala lie Met Gly 35 40 45

Phe He Gly Phe Phe Val Lys Leu He His He Pro He Asn Asn He

50 55 60

He Val Gly Gly 65

23

Claims

What is claimed is:

1. An isolated polynucleotide comprising a nucleotide sequence that has at least 80% identity over its entire length to a nucleotide sequence encoding the CBDAME07 polypeptide of SEQ ID NOJ; or a nucleotide sequence complementary to said isolated polynucleotide.

2. The polynucleotide of claim 1 wherein said polynucleotide comprises the nucleotide sequence contained in SEQ ID NOJ encoding the CBDAME07 polypeptide of SEQ ID N02.

3. The polynucleotide of claim 1 wherein said polynucleotide comprises a nucleotide sequence that is at least 80% identical to that of SEQ ID NO: 1 over its entire length.

4. The polynucleotide of claim 3 which is polynucleotide of SEQ ID NO: 1.

5. The polynucleotide of claim 1 which is DNA or RNA.

6. A DNA or RNA molecule comprising an expression system, wherein said expression system is capable of producing a CBDAME07 polypeptide comprising an amino acid sequence, of SEQ ID NOJ when said expression system is present in a compatible host cell.

7. A host cell comprising the expression system of claim 6.

8. A process for producing a CBDAME07 polypeptide comprising culturing a host of claim 7 under conditions sufficient for the production of said polypeptide and recovering the polypeptide from the culture.

9. A process for producing a cell which produces a CBDAME07 polypeptide thereof comprising transforming or transfecting a host cell with the expression system of claim 6 such that the host cell, under appropriate culture conditions, produces a CBDAME07 polypeptide.

10. A CBDAME07 polypeptide comprising an amino acid sequence of SEQ ID NOJ.

24

11 The polypeptide of claim 10 which compnses the ammo acid sequence of SEQ ID O 2

12 An antibody lmmunospecific for the CBDAME07 polypeptide of claim 10

13 A method for the treatment of a subject m need of enhanced activity or expression of CBDAME07 polypeptide of claim 10 compnsing

(a) administering to the subject a therapeutically effective amount of an agonist to said polypeptide, and/or

(b) providmg to the subject an isolated polynucleotide compnsing a nucleotide sequence that has at least 80% identity to a nucleotide sequence encodmg the CBDAME07 polypeptide of SEQ ID NO 2 over its entire length, or a nucleotide sequence complementary to said nucleotide sequence m a form so as to effect production of said polypeptide activity in vivo

14 A method for the treatment of a subject havmg need to inhibit activity or expression of CBDAME07 polypeptide of claim 10 compnsmg

(a) administering to the subject a therapeutically effective amount of an antagonist to said polypeptide, and/or (b) administering to the subject a nucleic acid molecule that inhibits the expression of the nucleotide sequence encodmg said polypeptide, and/or

(c) administenng to the subject a therapeutically effective amount of a polypeptide that competes with said polypeptide for its hgand, substrate , or receptor

15 A process for diagnosing a disease or a susceptibility to a disease in a subject related to expression or activity of CBDAME07 polypeptide of claim 10 m a subject compnsmg

(a) determining the presence or absence of a mutation m the nucleotide sequence encodmg said CBDAME07 polypeptide in the genome of said subject, and/or

(b) analyzing for the presence or amount of the CBDAME07 polypeptide expression m a sample derived from said subject

16 A method for identifying compounds which inhibit (antagonize) or agonize the

CBDAME07 polypeptide of claim 10 which comprises

25 (a) contacting a candidate compound with cells which express the CBDAME07 polypeptide (or cell membrane expressing CBDAME07 polypeptide) or respond to CBDAME07 polypeptide; and

(b) observing the binding, or stimulation or inhibition of a functional response; or comparing the ability of the cells (or cell membrane) which were contacted with the candidate compounds with the same cells which were not contacted for CBDAME07 polypeptide activity.

17. An agonist identified by the method of claim 16.

18. An antagonist identified by the method of claim 16.

19. A recombinant host cell produced by a method of Claim 9 or a membrane thereof expressing a CBDAME07 polypeptide.

26