HUMAN RECEPTOR GPR14, AND A METHOD OF FINDING AGONIST AND ANTAGONIST TO HUMAN AND RAT GPR14

Abstract: Human GPR14 polypeptides and polynucleotides and methods for producing such polypeptides by recombinant techniques are disclosed. Also disclosed are methods for utilizing Human GPR14 polypeptides and polynucleotides in the design of protocols for the treatment of ischemic coronary artery disease (angina and myocardial infarction); atherosclerosis; metabolic diseases (e.g. diabetes); CHF/myocardial dysfunction; arrhythmias; restenosis; hypertension; hypotension; pulmonary disease (hypertension, COPD, asthma); fibrotic vasculopathies (diabetes, SLE, AS, Reynaud's); cerebrovascular events (e.g. hemnorrhagic and ischemic stroke); neurogenic inflammation/migraine; hematopoietic disorders; ARDS; cancer; autoimmune diseases (e.g. HIV-1 and -2 infection and AIDS); gastrointestinal and genitourinary disturbances (e.g. ulcers); endocrine disorders; fibroproliferative disorders (e.g. psoriasis); inflammatory disease (e.g. RA, Crohn's, IBS); benign prostatic hypertrophy; renal failure and glomerulopathies; disease states, both cardiovascular and non-cardiovascular, which are characterized by excessive vasoconstriction, myocardial dysfunction and/or aberrant fibroproliferative/inflammatory responses; psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, delirium, dementia, severe mental retardation, Parkinson's disease, and dyskinesias, infections such as bacterial, fungal, protozoan and viral infections; pain; eating disorders, such as obesity, anorexia, and bulimia; asthma; urinary retention; oteoporosis; allergies; Huntington's disease or Gilles de la Tourette's syndrome, among others and diagnostic assays for such conditions.

Description: - 1 - HUMAN RECEPTOR GPR14, AND A METHOD OF FINDING AGONIST AND ANTAGONIST TO HUMAN AND RAT GPR14 This application claims the benefit of priority from U.S. Provisional Application Serial No. 60/074, 075 filed February 9, 1998 and U.S. Continuation-In-Part Application Serial No. to be provided filed January 15, 1999, which, in turn, claims priority of Continuation-In-Part Application Serial No. 09/058,725 filed April 10, 1998. All three applications are incorporated herein by reference in their entireties. 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 G-Protein coupled receptor, hereinafter referred to as Human GPR14. The invention also relates to inhibiting or activating the action of such polynucleotides and polypeptides. BACKGROUND OF THE INVENTION It is well established that many medically significant biological processes are mediated by proteins participating in signal transduction pathways that involve G-proteins and/or second messengers, e.g., cAMP (Lef owitz, Nature, 1991, 351:353-354). Herein these proteins are referred to as proteins participating in pathways with G-proteins or PPG proteins. Some examples of these proteins include the GPC receptors, such as those for adrenergic agents and dopamine (Kobilka, B.K., et al., Proc. Natl Acad. Sci., USA, 1987, 84:46-50; Kobilka, B.K., et al., Science, 1987, 238:650-656; Bunzow, J.R., et al., Nature, 1988, 336:783-787), G-proteins themselves, effector proteins, e.g., phospholipase C, adenyl cyclase, and phosphodiesterase, and actuator proteins, e.g., protein kinase A and protein kinase C (Simon, M.I., et al., Science, 1991, 252:802-8). For example, in one form of signal transduction, the effect of hormone binding is activation of the enzyme, adenylate cyclase, inside the cell. Enzyme activation by hormones is dependent on the presence of the nucleotide, GTP. GTP also influences hormone binding. A G-protein connects the hormone receptor to adenylate cyclase. G-protein was shown to exchange GTP for bound GDP when activated by a hormone receptor. The GTP-carrying - 2 - form then binds to activated adenylate cyclase. Hydrolysis of GTP to GDP, catalyzed by the G-protein itself, returns the G-protein to its basal, inactive form. Thus, the G-protein serves a dual role, as an intermediate that relays the signal from receptor to effector, and as a clock that controls the duration of the signal. The membrane protein gene superfamily of G-protein coupled receptors has been characterized as having seven putative transmembrane domains. The domains are believed to represent transmembrane a-helices connected by extracellular or cytoplasmic loops. G-protein coupled receptors include a wide range of biologically active receptors, such as hormone, viral, growth factor and neuroreceptors. G-protein coupled receptors have been characterized as including these seven conserved hydrophobic stretches of about 20 to 30 amino acids, connecting at least eight divergent hydrophilic loops. The G-protein family of coupled receptors includes dopamine receptors which bind to neuroleptic drugs used for treating psychotic and neurological disorders. Other examples of members of this family include, but are not limited to, calcitonin, adrenergic, endothelin, cAMP, adenosine, muscarinic, acetylcholine, serotonin, histamine, thrombin, kinin, follicle stimulating hormone, opsins, endothelial differentiation gene-1, rhodopsins, odorant, and cytomegaloviius receptors. Most G-protein coupled receptors (or otherwise known as 7TM receptors) have single conserved cysteine residues in each of the first two extracellular loops which form disulfide bonds that are believed to stabilize functional protein structure. The 7 transmembrane regions are designated as TM1, TM2, TM3, TM4, TM5, TM6, and TM7. TM3 has been implicated in signal transduction. Phosphorylation and lipidation (palmitylation or farnesylation) of cysteine residues can influence signal transduction of some G-protein coupled receptors. Most G-protein coupled receptors contain potential phosphorylation sites within the third cytoplasmic loop and/or the caiboxy terminus. For several G-protein coupled receptors, such as the β-adrenoreceptor, phosphorylation by protein kinase A and/or specific receptor kinases mediates receptor desensitization. For some receptors, the ligand binding sites of G-protein coupled receptors are believed to comprise hydrophilic sockets formed by several G-protein coupled receptor transmembrane domains, said sockets being surrounded by hydrophobic residues of the G- protein coupled receptors. The hydrophilic side of each G-protein coupled receptor transmembrane helix is postulated to face inward and form a polar ligand binding site. TM3 has been implicated in several G-protein coupled receptors as having a ligand binding site, such as the TM3 aspartate residue. TM5 serines, a TM6 asparagine and TM6 or TM7 phenylalanines or tyrosines are also implicated in ligand binding. G-protein coupled receptors can be intracellularly coupled by heterotrimeric G- proteins to various intracellular enzymes, ion channels and transporters (see, Johnson et al., Endoc. Rev., 1989, 10:317-331). Different G-protein a-subunits preferentially stimulate particular effectors to modulate various biological functions in a cell. Phosphorylation of cytoplasmic residues of G-protein coupled receptors has been identified as an important mechanism for the regulation of G-protein coupling of some G-protein coupled receptors. G-protein coupled receptors are found in numerous sites within a mammalian host. Over the past 15 years, nearly 350 therapeutic agents targeting 7 transmembrane (7 TM) receptors have been successfully introduced onto the market. This indicates that these receptors have an established, proven history as therapeutic targets. Clearly there is a need for identification and characterization of further receptors which can play a role in preventing, ameliorating or correcting dysfunctions or diseases, including, but not limited to, ischemic coronary artery disease (angina and myocardial infarction); atherosclerosis; metabolic diseases (e.g. diabetes); CHF/myocardial dysfunction; arrhythmias; restenosis; hypertension; hypotension; pulmonary disease (hypertension, COPD, asthma); fibrotic vasculopathies (diabetes, SLE, AS, Reynaud's); cerebrovascular events (e.g. hemnorrhagic and ischemic stroke); neurogenic inflammation/migraine; hematopoietic disorders; ARDS; cancer; autoimmune diseases (e.g. HTV-1 and -2 infection and AIDS); gastrointestinal and genitourinary disturbances (e.g. ulcers); endocrine disorders; fibroproliferative disorders (e.g. psoriasis); inflammatory disease (e.g. RA, Crohn's, IBS); benign prostatic hypertrophy; renal failure and glomerulopathies; disease states, both cardiovascular and non-cardiovascular, which are characterized by excessive vasoconstriction, myocardial dysfunction and/or aberrant fibroproliferative/inflammatory - 4 - responses; psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, delirium, dementia, severe mental retardation, Parkinson's disease, and dyskinesias, infections such as bacterial, fungal, protozoan and viral infections; pain; eating disorders, such as obesity, anorexia, and bulimia; asthma; urinary retention; osteoporosis; allergies; Huntington's disease or Gilles dela Tourett's syndrome. SUMMARY OF THE INVENTION In one aspect, the invention relates to Human GPR14 polypeptides and recombinant materials and methods for their production. Another aspect of the invention relates to methods for using such Human GPR14 polypeptides and polynucleotides. Such uses include the treatment of ischemic coronary artery disease (angina and myocardial infarction); atherosclerosis; metabolic diseases (e.g. diabetes); CHF/myocardial dysfunction; arrhythmias; restenosis; hypertension; hypotension; pulmonary disease (hypertension, COPD, asthma); fibrotic vasculopathies (diabetes, SLE, AS, Reynaud's); cerebrovascular events (e.g. hemnorrhagic and ischemic stroke); neurogenic inflammation/migraine; hematopoietic disorders; ARDS; cancer; autoimmune diseases (e.g. HIV-1 and -2 infection and AIDS); gastrointestinal and genitourinary disturbances (e.g. ulcers); endocrine disorders; fibroproliferative disorders (e.g. psoriasis); inflammatory disease (e.g. RA, Crohn's, LBS); benign prostatic hypertrophy; renal failure and glomerulopathies; disease states, both cardiovascular and non-cardiovascular, which are characterized by excessive vasoconstriction, myocardial dysfunction and/or aberrant fibroproUferative/inflammatory responses; psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, delirium, dementia, severe mental retardation, Parkinson's disease, and dyskinesias, infections such as bacterial, fungal, protozoan and viral infections; pain; eating disorders, such as obesity, anorexia, and bulimia; asthma; urinary retention; osteoporosis; allergies; Huntington's disease or Gilles dela Tourett's syndrome, among others. In accordance with another aspect of the present invention there are provided methods of screening for compounds which bind to and activate (agonist) or inhibit activation (antagonist) of rat or Human GPR 14 polypeptides (receptors), and for their ligands. - 5 - In particular, the preferred method for identifying agonist or antagonist of a Human or rat GPR 14 polypeptide comprises: contacting a cell expressing on the surface thereof the polypeptide, said polypeptide being associated with a second component capable of providing a detectable signal in response to the binding of a compound to said polypeptide, with a compound to be screened under conditions to permit binding to the polypeptide; and determining whether the compound binds to and activates or inhibits the polypeptide by measuring the level of a signal generated from the interaction of the compound with the polypeptide. In a further preferred embodiment, the method further comprises conducting the identification of agonist or antagonist in the presence of labeled or unlabeled, fish or Human urotensin LL In another embodiment of the method for identifying agonist or antagonist of a human or rat GPR 14 polypeptide comprises: determining the inhibition of binding of a Ugand to cells which have the polypeptide on the surface thereof, or to cell membranes containing the polypeptide, in the presence of a candidate compound under conditions to permit binding to the polypeptide, and determining the amount of Ugand bound to the polypeptide, such that a compound capable of causing reduction of binding of a Ugand is an agonist or antagonist. Preferably, the Ugand is human or rat urotensin π. Yet more preferably human or rat urotensin π is labeled. Further the present invention relates to treating conditions associated with Human GPR14 imbalance with the identified compounds. Yet another aspect of the invention relates to diagnostic assays for detecting diseases associated with inappropriate Human GPR14 activity or levels. BRIEF DESCRIPTION OF THE DRAWINGS Figures 1, 1A and IB show the nucleotide and deduced amino acid sequence of Human GPR14. SEQ ID NOS: 1 and 2. Figures 2A and 2B show the amino acid sequence and polynucleotide sequence of rat GPR14 receptor. SEQ ID NO: 4 and 6. - 6 - Figure 3 shows concentration response curves for fish urotensin π against HEK 293 rat and Human GPR14 transients + G alpha- 16 using FLIPR. Figure 4 is the concentration-response-curves for human urotensin π for 3 clones of the Human GPR 14 stably transfected ceUs. DESCRIPTION OF THE INVENTION Definitions The following definitions are provided to faciUtate understanding of certain terms used frequently herein. "Human GPR14" refers generally to a polypeptide having the amino acid sequence set forth in SEQ ID NO:2, or an allelic variant thereof. "Fish urotensin LT" refers to polypeptide with amino acid sequence of Ala-Gly- Thr-Ala-Asp-Cys-Phe-Trp-Lys-Tyr-Cys-Val (SEQ ID NO:3), which is described by Pearson et al. , in Proc Natl Acad Sci U S A 1980 Aug;77(8):5021-5024. "Rat GPR14 receptor" or "rat GPR 14 receptor polypeptide" is a protein with GenBank Accession number U32673, and described by Marchese et al. , in Genomics 1995 Sep 20;29(2):335-344. Rat GPR14 receptor has amino acid sequence shown in Figure 2 (SEQ ID NO: 4). "Human urotensin II" is a protein with the amino acid sequence ETPDCFWKYCV (SEQ ID NO: 5). Our studies have now found that Human urotensin π of SEQ ID NO: 5 to be the most potent mammaUan vasoconstrictor identified to date and, in the intact primate, induces systemic vasoconstriction, myocardial contractile dysfunction and, ultimately, lethal arrhythmias (additional metabolic/endocrine dysfunction may also result from excessive activity of this system). "Receptor Activity" or "Biological Activity of the Receptor" refers to the metabolic or physiologic function of said Human GPR14 including similar activities or improved activities or these activities with decreased undesirable side- effects. Also included are antigenic and immunogenic activities of said Human GPR14. "Human GPR14 polypeptides" refers to polypeptides with amino acid sequences sufficiently similar to Human GPR14 sequences, preferably exhibiting at least one biological activity of the receptor. - 7 - "Human GPR14 gene" refers to a polynucleotide having the nucleotide sequence set forth in SEQ ID NO: 1 or allelic variants thereof and/or their complements. "Human GPR14 polynucleotides" refers to polynucleotides containing a nucleotide sequence which encodes a Human GPR14 polypeptide or fragment thereof, or a nucleotide sequence which has at least 75.9 % identity to a nucleotide sequence encoding the polypeptide of SEQ ID NO: 2 or the corresponding fragment thereof, or 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. "Antibodies" as used herein includes polyclonal and monoclonal antibodies, chimeric, single chain, and humanized antibodies, as well as Fab fragments, including the products of an Fab or other immunoglobulin expression Ubrary. "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 polyribonucleotide or polydeoxribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA. "Polynucleotides" include, without limitation single- and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions, single- and double- stranded RNA, and RNA that is mixture of single- and double- stranded regions, hybrid molecules comprising DNA and RNA that may be single- stranded or, more typically, double- stranded or a mixture of single- and double-stranded regions. In addition, "polynucleotide" refers to triple-stranded regions comprising RNA or DNA or both RNA and DNA. The term polynucleotide also includes DNAs or RNAs containing one or more modified bases and DNAs or RNAs with backbones modified for stability or for other reasons. "Modified" bases include, for example, tritylated bases and unusual bases such as inosine. A variety of modifications has been made to DNA and RNA; thus, - 8 - "polynucleotide" embraces chemically, enzymatically or metaboUcally modified forms of polynucleotides as typically found in nature, as well as the chemical forms of DNA and RNA characteristic of viruses and cells. "Polynucleotide" also embraces relatively short polynucleotides, often referred to as oligonucleotides. "Polypeptide" refers to any peptide or protein comprising two or more amino acids joined to each other by peptide bonds or modified peptide bonds, i.e., peptide isosteres. "Polypeptide" refers to both short chains, commonly referred to as peptides, oligopeptides or oligomers, and to longer chains, generally referred to as proteins. Polypeptides may contain amino acids other than the 20 gene-encoded amino acids. "Polypeptides" include amino acid sequences modified either by 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 cycUc, with or without branching. CycUc, branched and branched cycUc 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 phosphotidyUnositol, 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, - 9 - 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 Umited 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 more substitutions, additions, deletions in 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 allelic variant, or it may be a variant that is not known to occur naturally. Non- naturally occurring variants of polynucleotides and polypeptides may be made by mutagenesis techniques or by direct synthesis. "Identity" as known in the art, is a relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as determined by comparing 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 Umited to those described in (Computational Molecular Biology, Lesk, A.M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D.W., ed., Academic Press, - 10 - New York, 1993; Computer Analysis of Sequence Data, Part i, Griffin, A.M. , and Griffin, H.G., eds. , Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G. , Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J. , eds. , M Stockton Press, New York, 1991; and Carillo, H., and Lipman, D., SIAM J. Applied Math. , 48: 1073 (1988). Preferred methods to determine identity are designed to give the largest match between the sequences tested. Methods to determine identity and similarity are codified in publicly available computer programs. Preferred computer program methods to determine identity and similarity between two sequences include, 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 NIH Bethesda, MD 20894; Altschul, S. , et al , J. Mol. Biol. 215: 403-410 (1990). The well known Smith Waterman algorithm may also be used to determine identity. Preferred parameters for polypeptide sequence comparison include the following: 1) Algorithm: Needleman and Wunsch, J. Mol Biol. 48: 443-453 (1970) Comparison 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 pubUcly available as the "gap" program from Genetics Computer Group, Madison WI. The aforementioned parameters are the default parameters for peptide comparisons (along with no penalty for end gaps). Preferred parameters for polynucleotide comparison include the following: 1) Algorithm: Needleman and Wunsch, J. Mol Biol. 48: 443-453 (1970) Comparison matrix: matches = +10, mismatch = 0 Gap Penalty: 50 - 11 - Gap Length Penalty: 3 Available as: The "gap" program from Genetics Computer Group, Madison WI. These are the default parameters for nucleic acid comparisons. Preferred polynucleotide embodiments further include an isolated polynucleotide comprising a polynucleotide sequence having at least a 50, 60, 70, 80, 85, 90, 95, 97 or 100% identity to the reference sequence of SEQ ID NO. l , wherein said polynucleotide sequence may be identical to the reference sequence of SEQ ID NO: 1 or may include 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, interspersed either individually among the nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence, and wherein said number of nucleotide alterations is determined by multiplying the total number of nucleotides in SEQ ID NO:l by the integer defining the percent identity divided by 100 and then subtracting that product from said total number of nucleotides in SEQ ID NO: l , or: n n x n - (x n y), wherein n n is the number of nucleotide alterations, x n is the total number of nucleotides in SEQ ID NO: l , 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 is the symbol for the multiplication operator, and wherein any non-integer product of x n and y is rounded down to the nearest integer prior to subtracting it from x n . 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 foUowing such alterations. By way of example, a polynucleotide sequence of the present invention may be identical to the reference sequence of SEQ ID NO:2, that is it may be 100% identical, or it may include up to a certain integer number of amino acid alterations as compared to the reference sequence such that the percent identity is less than 100% identity. Such alterations are selected from the group consisting of at least one nucleic acid deletion, substitution, , including transition and transversion, or insertion, and wherein said - 12 - alterations may occur at the 5' or 3' terminal positions of the reference polynucleotide sequence or anywhere between those terminal positions, interspersed either individually among the nucleic acids in the reference sequence or in one or more contiguous groups within the reference sequence. The number of nucleic acid alterations for a given percent identity is determined by multiplying the total number of amino acids in SEQ ID NO: 2 by the integer defining the percent identity divided by 100 and then subtracting that product from said total number of amino acids in SEQ ID NO: 2, or: n n = x n - (x n y), wherein n n is the number of amino acid alterations, x n is the total number of amino acids in SEQ ID NO:2, y is, for instance 0.70 for 70%, 0.80 for 80% , 0.85 for 85 % etc., , is the symbol for the multipUcation operator, and wherein any non-integer product of x n and y is rounded down to the nearest integer prior to subtracting it from x n . 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 polypeptide sequence may be identical to the reference 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 integer defining the percent identity divided by 100 and then subtracting that product from said total number of amino acids in SEQ ID NO: 2, or: n a x a - (x a y), wherein n a is the number of amino acid alterations, x a is the total number of amino acids in SEQ ID NO:2, 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 is the symbol for the multipUcation operator, and wherein any non- integer product of x a and y is rounded down to the nearest integer prior to subtracting it from x a . - 13 - By way of example, a polypeptide sequence of the present invention may be identical to the reference sequence of SEQ ID NO:2, that is it may be 100% identical, or it may include up to a certain integer number of amino acid alterations as compared to the reference sequence such that the percent identity is less than 100% identity. Such 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. The number of amino acid alterations for a given % identity is determined by multiplying the total number of amino acids in SEQ ID NO: 2 by the integer defining the percent identity divided by 100 and then subtracting that product from said total number of amino acids in SEQ ID NO:2, or: n a = x a - (x a y), wherein n a is the number of amino acid alterations, x a is the total number of amino acids in SEQ ID NO:2, y is, for instance 0.70 for 70% , 0.80 for 80% , 0.85 for 85 % etc. , and is the symbol for the multiplication operator, and wherein any non-integer product of x a and y is rounded down to the nearest integer prior to subtracting it from x a . Polypeptides of the Invention The Human GPR14 polypeptides of the present invention include the polypeptide of SEQ ID NO: 2 (in particular the mature polypeptide) as weU as Human GPR14 polypeptides and which have at least 84.25 % identity to the polypeptide of SEQ ID NO:2 or the relevant portion and more preferably at least 85% identity, and still more preferably at least 90% identity, and even still more preferably at least 95 % identity to SEQ ID NO: 2. The Human GPR14 polypeptides may be in the form of the "mature" protein or may be a part of a larger protein such as a fusion protein. It is often advantageous to include an additional amino acid sequence which contains secretory or leader sequences, pro-sequences, sequences which aid in purification such as multiple histidine residues, or an additional sequence for stabiUty during recombinant production. - 14 - BiologicaUy active fragments of the Human GPR14 polypeptides are also included in the invention. A fragment is a polypeptide having an amino acid sequence that entirely is the same as part, but not aU, of the amino acid sequence of the aforementioned Human GPR14 polypeptides. As with Human GPR14 polypeptides, fragments may be "free- standing, " or comprised 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 amino acid number 1-20, 21-40, 41-60, 61-80, 81-100, and 101 to the end of Human GPR14 polypeptide. In this context "about" includes the particularly recited ranges larger or s aUer by several, 5, 4, 3, 2 or 1 amino acid at either extreme or at both extremes. Preferred fragments include, for example, truncation polypeptides having the amino acid sequence of Human GPR14 polypeptides, except for deletion of a continuous series of residues that includes the amino terminus, or a continuous series of residues that includes the carboxyl terminus or deletion of two continuous series of residues, one including the amino terminus and one including the carboxyl terminus. Also preferred are fragments characterized by structural or functional attributes such as fragments that comprise alpha- heUx and alpha-helix forming regions, beta-sheet and beta-sheet-forming regions, turn and turn-forming regions, coU and coil-forming regions, hydrophiUc regions, hydrophobic regions, alpha amphipathic regions, beta amphipathic regions, flexible regions, surface- forming regions, substrate binding region, and high antigenic index regions. Biologically active fragments are those that mediate receptor activity, including those with a similar activity or an improved activity, or with a decreased undesirable activity. Also included are those that are antigenic or immunogenic in an animal, especially in a human. Thus, the polypeptides of the invention include polypeptides having an amino acid sequence at least 84.25 % identical to that of SEQ ID NO:2 or fragments thereof with at least 84.25 % identity to the corresponding fragment of SEQ ID NO:2. Preferably, aU of these polypeptides retain the biological activity of the receptor, including antigenic activity. Included in this group are variants of the defined sequence and fragments. Preferred variants are those that vary from the referents by conservative amino acid substitutions - i.e. , those that substitute a residue with another of like characteristics. Typical such substitutions are among Ala, Val, Leu and De; among Ser and Thr; among the acidic - 15 - residues Asp and Glu; among Asn and Gin; and among the basic residues Lys and Arg: or aromatic residues Phe and Tyr. 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 Human GPR14 polypeptides of the invention can be prepared in any suitable manner. Such polypeptides include isolated naturaUy occurring polypeptides, recombinantly produced polypeptides, syntheticaUy produced polypeptides, or polypeptides produced by a combination of these methods. Means for preparing such polypeptides are weU understood in the art. Polynucleotides of the Invention Another aspect of the invention relates to isolated polynucleotides which encode the Human GPR14 polypeptides and polynucleotides closely related thereto. Human GPR14 of the invention is structurally related to other proteins of the G- Protein coupled receptor, as shown by the results of sequencing the cDNA encoding Human GPR14. The cDNA sequence contains an open reading frame encoding a protein of 390 amino acids. Human GPR14 of Figure 1 (SEQ ED NO:2) has about 84.25 % identity (using Fasta) in 387 amino acid residues with Rattus norvegicus GPR14 orphan receptor ( A. Marches, et al., Genomics 29(2):335-344, 1995). Furthermore, human GPR14 (SEQ. ID NO:2) is 31.7% identical to human Somatostatin-3 receptor over 331 amino acid residues ( Y. Yamada et al., Mol. Endocrinol. 6(12):2136-2142 , 1992). Human GPR14 gene of Figure 1 (SEQ ID NO: 1) has about 75.9 % identity (using blast) in 1539 bp nucleotide residues with Rattus norvegicus GPR14 orphan receptor ( A. Marches, et al., Genomics 29(2): 335-344, 1995). Furthermore, human GGPR14(SEQ. ID NO: l) is 79% identical over 783bp to Rattus norvegicus G- protein coupled receptor SENR (M. Tal, et al. , Biochem. Biophys. Res. Commun. 209(2): 752-759 1995.) One polynucleotide of the present invention encoding Human GPR 14 may be obtained using standard cloning and screening, from a cDNA Ubrary derived from mRNA in cells of human placenta using the expressed sequence tag (EST) analysis (Adams, M.D. , et al. Science (1991) 252: 1651-1656; Adams, M.D. et al , Nature, (1992) 555:632-634; Adams, M.D., et al , Nature (1995) 377 Supp:3-174). Polynucleotides of the invention - 16 - can also be obtained from natural sources such as genomic DNA Ubraries or can be synthesized using well known and commercially available techniques. Thus, the nucleotide sequence encoding Human GPR14 polypeptides may be identical over its entire length to the coding sequence in Figure 1 (SEQ ID NO: l), or may be a degenerate form of this nucleotide sequence encoding the polypeptide of SEQ ID NO: 2, or may be highly identical to a nucleotide sequence that encodes the polypeptide of SEQ ID NO:2. Preferably, the polynucleotides of the invention contain a nucleotide sequence that is highly identical, at least 75.9% identical, with a nucleotide sequence encoding a Human GPR14 polypeptide, or at least 75.9% identical with the encoding nucleotide sequence set forth in Figure 1 (SEQ ID NO:l), or at least 75.9 % identical to a nucleotide sequence encoding the polypeptide of SEQ ID NO: 2. When the polynucleotides of the invention are used for the recombinant production of Human GPR14 polypeptide, the polynucleotide may include the coding sequence for the mature polypeptide or a fragment thereof, by itself; the coding sequence for the mature polypeptide or fragment in reading frame with other coding sequences, such as those encoding a leader or secretory sequence, a pre-, or pro- or prepro- protein sequence, or other fusion peptide portions. For example, a marker sequence which faciUtates purification 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 in the pQE vector (Qiagen, Inc.) and described in Gentz et al. , Proc Natl Acad Sci USA (1989) 86:821-824, or is an HA tag. The polynucleotide may also contain non-coding 5' and 3' sequences, such as transcribed, non-translated sequences, spUcing and polyadenylation signals, ribosome binding sites and sequences that stabUize mRNA. Among particularly preferred embodiments of the invention are polynucleotides encoding Human GPR14 polypeptides having the amino acid sequence of set out in Figure 1 (SEQ ID NO: 2) and variants thereof. Further preferred embodiments are polynucleotides encoding Human GPR14 variants that have the amino acid sequence of the Human GPR14 of Figure 1 (SEQ ID NO:2) in which several, 5-10, 1-5, 1-3, 1-2 or 1 amino acid residues are substituted, deleted or added, in any combination. - 17 - Further preferred embodiments of the invention are polynucleotides that are at least 75.9 % identical over their entire length to a polynucleotide encoding the Human GPR14 polypeptide having the amino acid sequence set out in Figure 1 (SEQ ID NO:2), and polynucleotides which are complementary to such polynucleotides. In this regard, polynucleotides at least 80 % identical over their entire length to the same are particularly preferred, and those with at least 90% are especiaUy 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. The present invention further relates to polynucleotides that hybridize to the herein above-described sequences. In this regard, the present invention especiaUy relates to polynucleotides which hybridize under stringent conditions to the herein above-described polynucleotides. As herein used, the term "stringent conditions" means hybridization wiU occur only if there is at least 95 % and preferably at least 97% identity between the sequences. Polynucleotides of the invention, which are sufficiently identical to a nucleotide sequence contained in SEQ ID NO:l, may be used as hybridization probes for cDNA and genomic DNA, to isolate fuU-length cDNAs and genomic clones encoding Human GPR14 and to isolate cDNA and genomic clones of other genes that have a high sequence similarity to the Human GPR14 gene. Such hybridization techniques are known to those of skiU in the art. Typically these nucleotide sequences are 70% identical, preferably 80% identical, more preferably 90% identical to that of the referent. The probes generaUy wUl comprise at least 15 nucleotides. Preferably, such probes wiU have at least 30 nucleotides and may have at least 50 nucleotides. Particularly preferred probes will range between 30 and 50 nucleotides. 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 CeUs, Expression The present invention also relates to vectors which comprise a polynucleotide or polynucleotides of the present invention, and host ceUs which are geneticaUy engineered - 18 - 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 ceUs 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 al. , 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 Upid-mediated transfection, electroporation, transduction, scrape loading, baUistic introduction or infection. Representative examples of appropriate hosts include bacterial cells, such as streptococci, staphylococci, E. coli, Streptomyces and Bacillus subtilis cells; fungal cells, such as yeast cells and Aspergillus ceUs; insect ceUs such as Drosophila S2 and Spodoptera Sf9 ceUs; animal cells such as CHO, COS, HeLa, C127, 3T3, BHK, 293 and Bowes melanoma ceUs; and plant ceUs. A great variety of expression systems can be used. Such systems include, among others, chromosomal, episomal and virus-derived systems, e.g., vectors derived from bacterial plasmids, from bacteriophage, 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 retroviruses, and vectors derived from combinations thereof, such as those derived from plasmid and bacteriophage genetic elements, such as cosmids and phagemids. The expression systems may contain control regions that regulate as weU as engender expression. GeneraUy, any system or vector suitable to maintain, propagate or express polynucleotides to produce a polypeptide in a host may be used. The appropriate nucleotide sequence may be inserted into an expression system by any of a variety of weU- known and routine techniques, such as, for example, those set forth in Sambrook et al , MOLECULAR CLONING, A LABORATORY MANUAL (supra). - 19 - For secretion of the translated protein into the lumen of the endoplasmic reticulum, into the periplasmic space or into the extraceUular environment, appropriate 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 Human GPR14 polypeptide is to be expressed for use in screening assays, generaUy, 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 in the screening assay. If Human GPR14 polypeptide is secreted into the medium, the medium can be recovered in order to recover and purify the polypeptide; if produced intracellularly, the cells must first be lysed before the polypeptide is recovered. Human GPR14 polypeptides can be recovered and purified from recombinant ceU cultures by weU-known methods including 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 Uquid chromatography is employed for purification. WeU known techniques for refolding proteins may be employed to regenerate active conformation when the polypeptide is denatured during isolation and or purification. Diagnostic Assays This invention also relates to the use of Human GPR14 polynucleotides for use as diagnostic reagents. Detection of a mutated form of Human GPR14 gene associated with a dysfunction wUl 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 Human GPR14. Individuals carrying mutations in the Human GPR14 gene may be detected at the DNA level by a variety of techniques. Nucleic acids for diagnosis may be obtained from a subject's ceUs, such as from blood, urine, saliva, tissue biopsy or autopsy material. The genomic DNA may be used directly for detection or may be amplified enzymaticaUy by using PCR or other amplification techniques prior to analysis. RNA or cDNA may also be used in sirmlar fashion. Deletions and insertions can be detected by a change in size of the amplified - 20 - product in comparison to the normal genotype. Point mutations can be identified by hybridizing amplified DNA to labeled Human GPR14 nucleotide sequences. Perfectly matched sequences can be distinguished from mismatched duplexes by RNase digestion or by differences in melting temperatures. DNA sequence differences may also be detected by alterations in electrophoretic mobiUty of DNA fragments in gels, with or without denaturing agents, or by direct DNA sequencing. 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 SI protection or the chemical cleavage method. See Cotton et al , Proc Natl Acad Sci USA (1985) 85: 4397-4401. The diagnostic assays offer a process for diagnosing or determining a susceptibiUty to ischemic coronary artery disease (angina and myocardial infarction); atherosclerosis; metaboUc diseases (e.g. diabetes); CHF/myocardial dysfunction; arrythmias; restenosis; hypertension; hypotension; pulmonary disease (hypertension, COPD, asthma); fibrotic vasculopathies (diabetes, SLE, AS, Reynaud's); cerebrovascular events (e.g. hemnorrhagic and ischemic stroke); neurogenic inflammation/migraine; hematopoietic disorders; ARDS; cancer; autoimmune diseases (e.g. HTV-l and -2 infection and AIDS); gastrointestinal and genitourinary disturbances (e.g. ulcers) ; endocrine disorders; fibroproUferative disorders (e.g. psoriasis); inflammatory disease (e.g. RA, Crohn's, IBS); benign prostatic hypertrophy; renal faUure and glomerulopathies; disease states, both cardiovascular and non- cardiovascular, which are characterized by excessive vasoconstriction, myocardial dysfunction and/or aberrant fibroproUferative/inflammatoiy responses; psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, delirium, dementia, severe mental retardation, Parkinson's disease, and dyskinesias, infections such as bacterial, fungal, protozoan and viral infections; pain; eating disorders, such as obesity, anorexia, and bulimia; asthma; urinary retention; osteoporosis; aUergies; Huntington's disease or GiUes dela Tourett's syndrome through detection of mutation in the Human GPR14 gene by the methods described. In addition, ischemic coronary artery disease (angina and myocardial infarction); atherosclerosis; metaboUc diseases (e.g. diabetes); CHF/myocardial dysfunction; arrythmias; restenosis; hypertension; hypotension; pulmonary disease (hypertension, COPD, asthma); fibrotic vasculopathies (diabetes, SLE, AS, Reynaud's); cerebrovascular events (e.g. - 21 - hemnorrhagic and ischemic stroke); neurogenic inflammation/migraine; hematopoietic disorders; ARDS; cancer; autoimmune diseases (e.g. HTV-l and -2 infection and ALDS); gastrointestinal and genitourinary disturbances (e.g. ulcers); endocrine disorders; fibroproliferative disorders (e.g. psoriasis); inflammatory disease (e.g. RA, Crohn's, IBS); benign prostatic hypertrophy; renal failure and glomerulopathies; disease states, both cardiovascular and non-cardiovascular, which are characterized by excessive vasoconstriction, myocardial dysfunction and/or aberrant fibroproUferative/inflammatory responses; psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, delirium, dementia, severe mental retardation, Parkinson's disease, and dyskinesias, infections such as bacterial, fungal, protozoan and viral infections; pain; eating disorders, such as obesity, anorexia, and bulimia; asthma; urinary retention; osteoporosis; aUergies; Huntington's disease or Gilles dela Tourett's syndrome, can be diagnosed by methods comprising determining from a sample derived from a subject an abnormally decreased or increased level of Human GPR14 polypeptide or Human GPR14 mRNA. Decreased or increased expression can be measured at the RNA level using any of the methods well known in the art for the quantitation of polynucleotides, such as, for example, PCR, RT-PCR, RNase protection, Northern blotting and other hybridization methods. Assay techniques that can be used to determine levels of a protein, such as an Human GPR14, in a sample derived from a host are weU-known to those of skiU in the art. Such assay methods include radioimmunoassays, competitive-binding assays, Western Blot analysis and ELISA assays. Chromosome Assays The nucleotide sequences of the present invention are also valuable for chromosome identification. The sequence is specificaUy targeted to and can hybridize with a particular location on an individual human chromosome. The mapping of relevant sequences to chromosomes according to the present invention is an important first step in 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, in V. McKusick, MendeUan Inheritance in Man (available on Une through Johns Hopkins University Welch Medical Library). The relationship between genes and diseases that have been mapped to - 22 - the same chromosomal region are then identified through Unkage analysis (coinheritance of physicaUy adjacent genes). The differences in the cDNA or genomic sequence between affected and unaffected individuals can also be determined. If a mutation is observed in some or aU of the affected individuals but not in any normal individuals, then the mutation is likely to be the causative agent of the disease. Antibodies The polypeptides of the invention or their fragments or analogs thereof, or ceUs expressing them can also be used as immunogens to produce antibodies immunospecific for the Human GPR14 polypeptides. The term "immunospecific" means that the antibodies have substantiaUy greater affinity for the polypeptides of the invention than their affinity for other related polypeptides in the prior art. Antibodies generated against the Human GPR14 polypeptides can be obtained by administering the polypeptides or epitope-bearing fragments, analogs or ceUs to an animal, preferably a nonhuman, using routine protocols. For preparation of monoclonal antibodies, any technique which provides antibodies produced by continuous ceU Une cultures can be used. Examples include the hybridoma technique (Kohler, G. and Milstein, C, Nature (1975) 256:495-497), the trioma technique, the human B-cell hybridoma technique (Kozbor et al. , Immunology Today (1983) 4:72) and the EBV-hybridoma technique (Cole et al , MONOCLONAL ANTIBODIES AND CANCER THERAPY, pp. 77-96, Alan R. Liss, Inc., 1985). Techniques for the production of single chain antibodies (U.S. Patent No. 4,946,778) can also be adapted to produce single chain antibodies to polypeptides of this invention. Also, transgenic mice, or other organisms including other mammals, may be used to express humanized antibodies. The above-described antibodies may be employed to isolate or to identify clones expressing the polypeptide or to purify the polypeptides by affinity chromatography. Antibodies against Human GPR14 polypeptides may also be employed to treat ischemic coronary artery disease (angina and myocardial infarction); atherosclerosis; metaboUc diseases (e.g. diabetes); CHF/myocardial dysfunction; arrythmias; restenosis; - 23 - hypertension; hypotension; pulmonary disease (hypertension, COPD, asthma); fibrotic vasculopathies (diabetes, SLE, AS, Reynaud's); cerebrovascular events (e.g. hemnorrhagic and ischemic stroke); neurogenic inflammation/migraine; hematopoietic disorders; ARDS; cancer; autoimmune diseases (e.g. HTV-l and -2 infection and AIDS); gastrointestinal and genitourinary disturbances (e.g. ulcers); endocrine disorders; fibroproUferative disorders (e.g. psoriasis); inflammatory disease (e.g. RA, Crohn's, ESS); benign prostatic hypertrophy; renal faUure and glomerulopathies; disease states, both cardiovascular and non- cardiovascular, which are characterized by excessive vasoconstriction, myocardial dysfunction and/or aberrant fibroproliferative/inflammatory responses; psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, deUrium, dementia, severe mental retardation, Parkinson's disease, and dyskinesias, infections such as bacterial, fungal, protozoan and viral infections; pain; eating disorders, such as obesity, anorexia, and bulimia; asthma; urinary retention; osteoporosis; aUergies; Huntington's disease or GiUes dela Tourett's syndrome, among others. Vaccines Another aspect of the invention relates to a method for inducing an immunological response in a mammal which comprises inoculating the mammal with Human GPR14 polypeptide, or a fragment thereof, adequate to produce antibody and/or T cell immune response to protect said animal from ischemic coronary artery disease (angina and myocardial infarction); atherosclerosis; metabolic diseases (e.g. diabetes); CHF/myocardial dysfunction; arrythmias; restenosis; hypertension; hypotension; pulmonary disease (hypertension, COPD, asthma); fibrotic vasculopathies (diabetes, SLE, AS, Reynaud's); cerebrovascular events (e.g. hemnorrhagic and ischemic stroke); neurogenic inflammation/migraine; hematopoietic disorders; ARDS; cancer; autoimmune diseases (e.g. HTV-l and -2 infection and AIDS); gastrointestinal and genitourinary disturbances (e.g. ulcers); endocrine disorders; fibroproUferative disorders (e.g. psoriasis); inflammatory disease (e.g. RA, Crohn's, IBS); benign prostatic hypertrophy; renal faUure and glomerulopathies; disease states, both cardiovascular and non-cardiovascular, which are characterized by excessive vasoconstriction, myocardial dysfunction and/or aberrant fibroproUferative/inflammatory responses; psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, deUrium, dementia, severe mental retardation, - 24 - Parkinson's disease, and dyskinesias, infections such as bacterial, fungal, protozoan and viral infections; pain; eating disorders, such as obesity, anorexia, and bulimia; asthma; urinary retention; osteoporosis; aUergies; Huntington's disease or GiUes dela Tourett's syndrome, among others. Yet another aspect of the invention relates to a method of inducing immunological response in a mammal which comprises, delivering Human GPR14 gene via a vector directing expression of Human GPR14 polypeptide in vivo in order to induce such an immunological response to produce antibody to protect said animal from diseases. Further aspect of the invention relates to an immunological/vaccine formulation (composition) which, when introduced into a mammalian host, induces an immunological response in that mammal to a Human GPR14 polypeptide wherein the composition comprises a Human GPR14 polypeptide or Human GPR14 gene. The vaccine formulation may further comprise a suitable carrier. Since Human GPR14 polypeptide may be broken down in the stomach, it is preferably administered parenterally (including subcutaneous, intramuscular, intravenous, intradermal etc. injection). Formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic 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 Uquid 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 wiU depend on the specific activity of the vaccine and can be readily determined by routine experimentation. Screening Assays A Human GPR14 polypeptide (receptor of the present invention) may be employed in a screening process for compounds which bind the receptor and which activate (agonists) or inhibit activation of (antagonists) the receptor polypeptide of the present invention. Thus, polypeptides of the invention may also be used to assess the binding of smaU molecule substrates and Ugands in, for example, ceUs, ceU-free preparations, chemical Ubraries, and - 25 - natural product mixtures. These substrates and Ugands may be natural substrates and ligands or may be structural or functional mimetics. See CoUgan et al. , Current Protocols in Immunology l(2):Chapter 5 (1991). In general, such screening procedures involve providing appropriate cells which express a receptor polypeptide of the present invention on the surface thereof. Such cells include cells from mammals, yeast, Drosophila or E. coli. In particular, a polynucleotide encoding the receptor of the present invention is employed to transfect ceUs to thereby express a Human GPR14 polypeptide. The expressed receptor is then contacted with a test compound to observe binding, stimulation or inhibition of a functional response. One such screening procedure involves the use of melanophores which are transfected to express a Human GPR14 polypeptide. Such a screening technique is described in PCT WO 92/01810, published February 6, 1992. Such an assay may be employed to screen for a compound which inhibits activation of a receptor of the present invention by contacting the melanophore ceUs which encode the receptor with both a receptor ligand, such as human or fish urotensin π, and a compound to be screened. Inhibition of the signal generated by the Ugand indicates that a compound is a potential antagonist for the receptor, . e. , inhibits activation of the receptor. The technique may also be employed for screening of compounds which activate a receptor of the present invention by contacting such ceUs with compounds to be screened and determining whether such compound generates a signal, . e. , activates the receptor. Other screening techniques include the use of ceUs which express a Human GPR14 polypeptide (for example, transfected CHO ceUs) in a system which measures extraceUular pH changes caused by receptor activation. In this technique, compounds may be contacted with cells expressing a receptor polypeptide of the present invention. A second messenger response, e.g. , signal transduction or pH changes, is then measured to determine whether the potential compound activates or inhibits the receptor. Another screening technique involves expressing a Human GPR14 polypeptide in which the receptor is linked to phosphoUpase C or D. Representative examples of such ceUs include, but are not Umited to, endotheUal ceUs, smooth muscle ceUs, and - 26 - embryonic kidney ceUs. The screening may be accomplished as hereinabove described by detecting activation of the receptor or inhibition of activation of the receptor from the phospholipase second signal. Another method involves screening for compounds which are antagonists, and thus inhibit activation of a receptor polypeptide of the present invention by determining inhibition of binding of labeled ligand, such as fish or human urotensin π, to cells which have the receptor on the surface thereof, or cell membranes containing the receptor. Such a method involves transfecting a eukaryotic cell with a DNA encoding a Human GPR14 polypeptide such that the ceU expresses the receptor on its surface. The ceU is then contacted with a potential antagonist in the presence of a labeled form of a ligand, such as fish or human urotensin JJ. The ligand can be labeled, e.g., by radioactivity. The amount of labeled Ugand bound to the receptors is measured, e.g. , by measuring radioactivity associated with transfected cells or membrane from these ceUs. If the compound binds to the receptor, the binding of labeled ligand to the receptor is inhibited as determined by a reduction of labeled Ugand which binds to the receptors. This method is called binding assay. Another such screening procedure involves the use of mammalian cells which are transfected to express a receptor of the present invention. The cells are loaded with an indicator dye that produces a fluorescent signal when bound to calcium, and the cells are contacted with a test substance and a receptor agonist, such as fish or human urotensin π. Any change in fluorescent signal is measured over a defined period of time using, for example, a fluorescence spectrophotometer or a fluorescence imaging plate reader. A change in the fluorescence signal pattern generated by the Ugand indicates that a compound is a potential antagonist (or agonist) for the receptor. Another such screening procedure involves use of mammaUan ceUs which are transfected to express a receptor of the present invention, and which are also transfected with a reporter gene construct that is coupled to activation of the receptor (for example, luciferase or beta-galactosidase behind an appropriate promoter). The cells are contacted with a test substance and a receptor agonist, such as fish or human urotensin π, and the signal produced by the reporter gene is measured after a defined period of time. The signal can be measured using a luminometer, spectrophotometer, fluorimeter, or other - 27 - such instrument appropriate for the specific reporter construct used. Inhibition of the signal generated by the ligand indicates that a compound is a potential antagonist for the receptor. Another such screening technique for antagonists or agonists involves introducing RNA encoding a Human GPR14 polypeptide into Xenopus oocytes to transiently or stably express the receptor. The receptor oocytes are then contacted with a receptor ligand, such as human or fish urotensin π, and a compound to be screened. Inhibition or activation of the receptor is then determined by detection of a signal, such as, cAMP, calcium, proton, or other ions. Another method involves screening for a Human GPR14 polypeptide inhibitors by determining inhibition or stimulation of Human GPR14 polypeptide-mediated cAMP and/or adenylate cyclase accumulation or dimunition. Such a method involves transiently or stably transfecting a eukaryotic cell with a Human GPR14 polypeptide to express the receptor on the cell surface. The ceU is then exposed to potential antagonists in the presence of Human GPR14 polypeptide ligand, such as fish or human urotensin π. The amount of cAMP accumulation is then measured, for example, by radio-immuno or protein binding assays (for example using Flashplates or a scintiUation proximity assay). Changes in cAMP levels can also be determined by directly measuring the activity of the enzyme, adenylyl cyclase, in broken ceU preparations. If the potential antagonist binds the receptor, and thus inhibits Human GPR14 polypeptide binding, the levels of Human GPR14 polypeptide-mediated cAMP, or adenylate cyclase activity, wiU be reduced or increased. Another screening method for agonists and antagonists reUes on the endogenous pheromone response pathway in the yeast, Saccharomyces cerevisiae. HeterothaUic strains of yeast can exist in two mitoticaUy stable haploid mating types, MATa and MATa. Each cell type secretes a smaU peptide hormone that binds to a G-protein coupled receptor on opposite mating-type ceUs which triggers a MAP kinase cascade leading to Gl arrest as a prelude to ceU fusion. Genetic alteration of certain genes in the pheromone response pathway can alter the normal response to pheromone, and heterologous expression and coupling of human G-protein coupled receptors and humanized G-protein subunits in yeast ceUs devoid of endogenous pheromone receptors can be tinked to downstream - 28 - signaling pathways and reporter genes (e.g. , U.S. Patents 5,063,154; 5,482,835; 5,691 , 188). Such genetic alterations include, but are not limited to, (i) deletion of the S7E2 or STE3 gene encoding the endogenous G-protein coupled pheromone receptors; (ii) deletion of the FAR1 gene encoding a protein that normaUy associates with cycUn- dependent kinases leading to cell cycle arrest; and (iii) construction of reporter genes fused to the FUS1 gene promoter (where FUS1 encodes a membrane-anchored glycoprotein required for ceU fusion). Downstream reporter genes can permit either a positive growth selection (e.g. , histidine prototrophy using the FUS1-HIS3 reporter), or a colorimetric, fluorimetric or spectrophotometric readout, depending on the specific reporter construct used (e.g. , ?-galactosidase induction using a FUSl-LacZ reporter). The yeast cells can be further engineered to express and secrete smaU peptides from random peptide libraries, some of which can permit autocrine activation of heterologously expressed human (or mammalian) G-protein coupled receptors (Broach, J.R. and Thomer, J. Nature 384: 14-16, 1996; Manfredi et al. , Mol. Cell. Biol. 16: 4700-4709, 1996). This provides a rapid direct growth selection (e.g. , using the FU SI -HI S3 reporter) for surrogate peptide agonists that activate characterized or orphan receptors. Alternatively, yeast cells that functionally express human (or mammaUan) G-protein coupled receptors linked to a reporter gene readout (e.g. , FUSl-LacZ) can be used as a platform for high- throughput screening of known ligands, fractions of biological extracts and libraries of chemical compounds for either natural or surrogate Ugands. Functional agonists of sufficient potency (whether natural or surrogate) can be used as screening tools in yeast ceU-based assays for identifying G-protein coupled receptor antagonists. For this purpose, the yeast system offers advantages over mammalian expression systems due to its ease of utility and null receptor background (lack of endogenous G-protein coupled receptors) which often interferes with the abitity to identify agonists or antagonists. The present invention also provides a method for determining whether a Ugand not known to be capable of binding to a Human GPR14 polypeptide can bind to such receptor which comprises contacting a mammaUan ceU which expresses a Human GPR14 polypeptide with the Ugand, such as fish or urotensin π, under conditions permitting - 29 - binding of candidate ligands to a Human GPR14 polypeptide, and detecting the presence of a candidate ligand which binds to the receptor thereby determining whether the ligand binds to the Human GPR14 polypeptide. The systems hereinabove described for determining agonists and/or antagonists may also be employed for determining Ugands which bind to the receptor. Applicants further discovered that fish or human urotensin π binds equally well to rat GPR14 receptor. Thus all the above described assay techniques can be used for finding agonists and antagonists to rat GPR 14 receptor by employing fish or human urotensinll as a ligand; furthermore, such identified agonists and antagonists are believed to be also agonists and antagonists to Human GPR14 polypeptides. Further aspect of present invention relates to a method of screening for agonists and antagonists of Human GPR14 polypeptides comprising screening for agonists and antagonist for rat GPR14, preferably in the presence of fish or human urotensin II . Examples of potential Human GPR14 polypeptide antagonists include antibodies or, in some cases, oUgonucleotides, which bind to the receptor but do not eUcit a second messenger response such that the activity of the receptor is prevented. Potential antagonists also include proteins which are closely related to a ligand of the Human GPR14 polypeptide, i.e. a fragment of the ligand, which have lost biological function and when binding to the Human GPR14 polypeptide, eUcit no response. Thus in another aspect, the present invention relates to a screening kit for identifying agonists, antagonists, and Ugands for Human GPR14 polypeptides, which comprises: (a) rat GPR14 or a Human GPR14 polypeptide, preferably that of SEQ ID NO:2, and preferably further comprises labeled or unlabeled fish or human urotensin II; (b) a recombinant cell expressing rat GPR14 or a Human GPR14 polypeptide, preferably that of SEQ ID NO:2, and preferably further comprises labeled or unlabeled fish or human urotensin II; or (c) a ceU membrane expressing rat GPR14 or a Human GPR14 polypeptide; preferably that of SEQ ID NO: 2, and preferably further comprises labeled or unlabeled fish or human urotensin π. - 30 - It will be appreciated that in any such kit, (a), (b), or (c) may comprise a substantial component. A potential antagonist also includes an antisense construct prepared through the use of antisense technology. Antisense technology can be used to control gene expression through triple-helix formation or antisense DNA or RNA, both methods of which are based on binding of a polynucleotide to DNA or RNA. For example, the 5' coding portion of the polynucleotide sequence, which encodes for the mature polypeptides of the present invention, is used to design an antisense RNA oUgonucleotide of from about 10 to 40 base pairs in length. A DNA oUgonucleotide is designed to be complementary to a region of the gene involved in transcription (triple helix -see Lee, et al. Nucl. Acids Res. , 6: 3073 (1979); Cooney, et al, Science, 2M: 456 (1988); and Dervan, et al , Science, 251: 1360 (1991)), thereby preventing transcription and production of a Human GPR14 polypeptide. The antisense RNA oUgonucleotide hybridizes to the mRNA in vivo and blocks translation of the mRNA molecule to a Human GPR14 polypeptide (antisense - Okano, J. , Neurochem. , 56: 560 (1991); OLIGODEOXYNUCLEOTIDES AS ANTISENSE INHIBITORS OF GENE EXPRESSION, CRC Press, Boca Raton, FL (1988)). The oUgonucleotides described above can also be deUvered to cells such that the antisense RNA or DNA may be expressed in vivo to inhibit production of a Human GPR14 polypeptide. Another potential antagonist is a smaU molecule which binds to a Human GPR14 polypeptide, making it inaccessible to Ugands such that normal biological activity is prevented. Examples of smaU molecules include, but are not limited to, small peptides or peptide-like molecules. Potential antagonists also include soluble forms of a Human GPR14 polypeptide, e.g. , fragments of the polypeptide, which bind to the ligand and prevent the Ugand from interacting with membrane bound Human GPR 14 polypeptides. Human GPR14 proteins are ubiquitous in the mammalian host and are responsible for many biological functions, including many pathologies. Accordingly, it is desirous to find compounds and drugs which stimulate Human GPR14 on the one hand and which can inhibit the function of Human GPR14 on the other hand. In general, agonists are - 31 - employed for therapeutic and prophylactic purposes for such conditions as ischemic coronary artery disease (angina and myocardial infarction); atherosclerosis; metabolic diseases (e.g. diabetes); CHF/myocardial dysfunction; arrythmias; restenosis; hypertension; hypotension; pulmonary disease (hypertension, COPD, asthma); fibrotic vasculopathies (diabetes, SLE, AS, Reynaud's); cerebrovascular events (e.g. hemnorrhagic and ischemic stroke); neurogenic inflammation/migraine; hematopoietic disorders; ARDS; cancer; autoimmune diseases (e.g. HIV-1 and -2 infection and AIDS); gastrointestinal and genitourinary disturbances (e.g. ulcers); endocrine disorders; fibroproUferative disorders (e.g. psoriasis); inflammatory disease (e.g. RA, Crohn's, EBS); benign prostatic hypertrophy; renal failure and glomerulopathies; disease states, both cardiovascular and non-cardiovascular, which are characterized by excessive vasoconstriction, myocardial dysfunction and/or aberrant fibroproUferative/inflammatory responses; psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, delirium, dementia, severe mental retardation, Parkinson's disease, and dyskinesias, infections such as bacterial, fungal, protozoan and viral infections; pain; eating disorders, such as obesity, anorexia, and bulimia; asthma; urinary retention; osteoporosis; allergies; Huntington's disease or GiUes dela Tourett's syndrome. Antagonists may be employed for a variety of therapeutic and prophylactic purposes for such conditions as ischemic coronary artery disease (angina and myocardial infarction); atherosclerosis; metabolic diseases (e.g. diabetes); CHF/myocardial dysfunction; arrythmias; restenosis; hypertension; hypotension; pulmonary disease (hypertension, COPD, asthma); fibrotic vasculopathies (diabetes, SLE, AS, Reynaud's); cerebrovascular events (e.g. hemnorrhagic and ischemic stroke); neurogenic inflammation/migraine; hematopoietic disorders; ARDS; cancer; autoimmune diseases (e.g. HTV-l and -2 infection and AIDS); gastrointestinal and genitourinary disturbances (e.g. ulcers); endocrine disorders; fibroproUferative disorders (e.g. psoriasis); inflammatory disease (e.g. RA, Crohn's, EBS); benign prostatic hypertrophy; renal failure and glomerulopathies; disease states, both cardiovascular and non-cardiovascular, which are characterized by excessive vasoconstriction, myocardial dysfunction and/or aberrant fibroproUferative/inflammatory responses; psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, deUrium, dementia, severe mental retardation, - 32 - Parkinson's disease, and dyskinesias, infections such as bacterial, fungal, protozoan and viral infections; pain; eating disorders, such as obesity, anorexia, and bulimia; asthma; urinary retention; osteoporosis; aUergies; Huntington's disease or Gilles dela Tourett's syndrome. Prophylactic and Therapeutic Methods This invention provides methods of treating an abnormal conditions related to both an excess of and insufficient amounts of Human GPR14 activity. If the activity of Human GPR14 is in excess, several approaches are avaUable. One approach comprises administering to a subject an inhibitor compound (antagonist) as hereinabove described along with a pharmaceuticaUy acceptable carrier in an amount effective to inhibit activation by blocking binding of Ugands to the Human GPR14, or by inhibiting a second signal, and thereby aUeviating the abnormal condition. In another approach, soluble forms of Human GPR14 polypeptides stiU capable of binding the ligand in competition with endogenous Human GPR14 may be administered. Typical embodiments of such competitors comprise fragments of the Human GPR14 polypeptide. In still another approach, expression of the gene encoding endogenous Human GPR14 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 OUgodeoxynucleotides as Antisense Inhibitors of Gene Expression, CRC Press, Boca Raton, FL (1988). Alternatively, oUgonucleotides which form triple heUces with the gene can be suppUed. See, for example, Lee et al , Nucleic Acids Res (1979) 6:3073; Cooney et al, Science (1988) 241 :456; Dervan et al, Science (1991) 251:1360. These oUgomers can be administered per se or the relevant oUgomers can be expressed in vivo. For treating abnormal conditions related to an under-expression of Human GPR14 and its activity, several approaches are also available. One approach comprises administering to a subject a therapeuticaUy effective amount of a compound which activates Human GPR14, i.e., an agonist as described above, in combination with a pharmaceuticaUy acceptable carrier, to thereby aUeviate the abnormal condition. Alternatively, gene therapy - 33 - may be employed to effect the endogenous production of Human GPR14 by the relevant cells in the subject. For example, a polynucleotide of the invention may be engineered for expression in a repUcation defective retroviral vector, as discussed above. The retroviral expression construct may then be isolated and introduced into a packaging ceU transduced with a retroviral plasmid vector containing RNA encoding a polypeptide of the present invention such that the packaging ceU now produces infectious viral particles containing the gene of interest. These producer cells may be administered to a subject for engineering ceUs 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 Approache , (and references cited therein) in Human Molecular Genetics, T Strachan and A P Read, BIOS Scientific Publishers Ltd (1996). Formulation and Administration Peptides, such as the soluble form of Human GPR14 polypeptides, and agonists and antagonist peptides or smaU molecules, may be formulated in combination with a suitable pharmaceutical carrier. Such formulations comprise a therapeuticaUy effective amount of the polypeptide or compound, and a pharmaceuticaUy 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 weU within the skiU of the art. The invention further relates to pharmaceutical packs and kits comprising one or more containers fiUed 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, typicaUy 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. - 34 - Administration of these compounds may also be topical and/or localized, in the form of salves, pastes, gels and the like. 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 0.1- 100 μg/kg of subject. Wide variations in the needed dosage, however, are to be expected in view of the variety of compounds avaUable 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 weU understood in the art. Polypeptides used in treatment can also be generated endogenously in the subject, in treatment modaUties often referred to as "gene therapy" as described above. Thus, for example, ceUs 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 ceUs are then introduced into the subject. Examples The examples below are carried out using standard techniques, which are weU known and routine to those of skUl in the art, except where otherwise described in detaU. The examples Ulustrate, but do not limit the invention. Example 1 A 1.2 kb PCR fragment corresponding to the entire coding region of the Rattus norvegicus orphan receptor (GPR14, A. Marchese et al. Genomics, 29(2): 335-44, 1995) was used as a probe to screen a total of .75M plaques from a Human Genomic Placenta Ubrary (Stratagene, LaJoUa CA, Cat. # 946206). The genomic Ubrary screening procedure is described by Elgin, et al. Stratatgies 4: 8-9, 1991. The probes were α -32P labeled, using Random Primed LabeUng Kit (Boheringer Manheim, Germany, Cat. # 1585584) and purified by running over Sephadex G-50 columns (Pharmacia Biotech. Cat. # 17-0855- 02). The hybridization and washing conditions were according to J. Sambrook, E.F. Fritch and T. Maniatis (1989) A Laboratory Manual Second. Ed. Vol. 1 pp. 2.69-2.81 Cold Spring - 35 - Harbor Laboratory Press, Cold Spring Harbor, New York). One Positive phage clone obtained from above screen was further purified. Southern analysis as carried out on digested phage DNA and a 9 kb Sac I fragment hybridized for this clone. This fragment was subcloned into pBlueScript KS vector, and further digested with EcoRI and southern analyzed. A smaUer 5kb fragment hybridized to the rat GPR14 probe mentioned above was subcloned into pBlueScript for sequence analysis, the sequence was determined by automated sequencer. A total of 2126 bp were sequenced, this includes an open reading frame encoding a peptide of 390 residues. This sequence is highly homologous to Rat GPR14, as shown by sequence comparisons and also by fasta analysis against the Genbank nucleotide data base. Example 2: MammaUan CeU Expression The receptors of the present invention are expressed in either human embryonic kidney 293 (HEK293) cells or adherent dhfr CHO ceUs. To maximize receptor expression, typicaUy all 5' and 3' untranslated regions (UTRs) are removed from the receptor cDNA prior to insertion into a pCDN or pCDNA3 vector. The ceUs are transfected with individual receptor cDNAs by Upofectin and selected in the presence of 400 mg/ml G418. After 3 weeks of selection, individual clones are picked and expanded for further analysis. HEK293 or CHO ceUs transfected with the vector alone serve as negative controls. To isolate ceU lines stably expressing the individual receptors, about 24 clones are typically selected and analyzed by Northern blot analysis. Receptor mRNAs are generally detectably in about 50% of the G418-resistant clones analyzed. Example 3: Ligand bank for binding and functional assays. A bank of over 200 putative receptor Ugands has been assembled for screening. The bank comprises: transmitters, hormones and chemokines known to act via a human seven transmembrane (7TM) receptor; naturaUy occurring compounds which may be putative agonists for a human 7TM receptor, non-mammaUan, biologicaUy active peptides for which a mammaUan counterpart has not yet been identified; and compounds not found in nature, but which activate 7TM receptors with unknown natural Ugands. This bank is used to initiaUy screen the receptor for known Ugands, using both functional (i.e. calcium, cAMP, microphysiometer, oocyte electrophysiology, etc, see below) as weU as binding assays. - 36 - Example 4: Ligand Binding Assays Ligand binding assays provide a direct method for ascertaining receptor pharmacology and are adaptable to a high throughput format. The purified Ugand for a receptor is radiolabeled to high specific activity (50-2000 Ci/mmol) for binding studies. A determination is then made that the process of radiolabeling does not diminish the activity of the Ugand towards its receptor. Assay conditions for buffers, ions, pH and other modulators such as nucleotides are optimized to estabUsh a workable signal to noise ratio for both membrane and whole ceU receptor sources. For these assays, specific receptor binding is defined as total associated radioactivity minus the radioactivity measured in the presence of an excess of unlabeled competing Ugand. Where possible, more than one competing Ugand is used to define residual nonspecific binding. Example 5: Functional Assay in Xenopus Oocytes Capped RNA transcripts from Unearized plasmid templates encoding the receptor cDNAs of the invention are synthesized in vitro with RNA polymerases in accordance with standard procedures. In vitro transcripts are suspended in water at a final concentration of 0.2 mg/ml. Ovarian lobes are removed from adult female toads, Stage V defoUiculated oocytes are obtained, and RNA transcripts (10 ng/oocyte) are injected in a 50 nl bolus using a microinjection apparatus. Two electrode voltage clamps are used to measure the currents from individual Xenopus oocytes in response to agonist exposure.. Recordings are made in Ca2+ free Barth's medium at room temperature. The Xenopus system can be used to screen known Ugands and tissue/ceU extracts for activating Ugands. Example 6: Microphysiometric Assays Activation of a wide variety of secondary messenger systems results in extrusion of smaU amounts of acid from a ceU. The acid formed is largely as a result of the increased metaboUc activity required to fuel the intraceUular signaling process. The pH changes in the media surrounding the ceU are very smaU but are detectable by the CYTOSENSOR microphysiometer (Molecular Devices Ltd., Menlo Park, CA). The CYTOSENSOR is thus capable of detecting the activation of a receptor which is coupled to an energy utilizing intraceUular signaling pathway such as the G-protein coupled receptor of the present invention. - 37 - Example 7: Extract/Cell Supernatant Screening A large number of mammalian receptors exist for which there remains, as yet, no cognate activating Ugand (agonist). Thus, active Ugands for these receptors may not be included within the Ugands banks as identified to date. Accordingly, the 7TM receptor of the invention is also functionaUy screened (using calcium, cAMP, microphysiometer, oocyte electrophysiology, etc., functional screens) against tissue extracts to identify natural Ugands. Extracts that produce positive functional responses can be sequentially subfractionated until an activating ligand is isolated and identified. Example 8: Calcium and cAMP Functional Assays 7TM receptors which are expressed in HEK 293 ceUs have been shown to be coupled functionaUy to activation of PLC and calcium mobilization and/or cAMP stimulation or inhibition. Basal calcium levels in the HEK 293 cells in receptor-transfected or vector control cells were observed to be in the normal, 100 nM to 200 nM, range. HEK 293 ceUs expressing recombinant receptors are loaded with fura 2 and in a single day > 150 selected Ugands or tissue/ceU extracts are evaluated for agonist induced calcium mobilization. SimUarly, HEK 293 ceUs expressing recombinant receptors are evaluated for the stimulation or inhibition of cAMP production using standard cAMP quantitation assays. Agonists presenting a calcium transient or cAMP fluctuation are tested in vector control ceUs to determine if the response is unique to the transfected ceUs expressing receptor. Example 9: Urotensin H-induced Ca2 + mobilization in transient HEK 293 GPR14 and stable HEK 293 G?16 GPR14 ceUs. The GPR14 receptor when transiently transfected into HEK 293 ceUs responds to urotensin π in a concentration-dependent manner with a robust calcium response. Both the transiently transfected rat and human receptors respond to fish urotensin π with similar affinities. Figure 3 shows the concentration response curves for fish urotensin π against both the rat and human receptors. The data was generated with the 96 weU FLuorescent Imaging Plate Reader (FLIPR). Each point is the mean of 6-8 individual weUs in 3-4 separate 96 weU plates read on FLIPR. The EC50 for fish urotensin II was 0.5 nM for the human receptor and 0.8 nM for the rat receptor. - 38 - AdditionaUy the human receptor was stably transfected into HEK 293 G?16 ceUs and human urotensin π produced concentration dependent calcium mobilization responses. Figure 4 is the concentration-response-curves for human urotensin π for 3 clones (designated L2, L3 and L4) of the stably transfected cells. The EC50s of human urotensin π for the 3 clones were 0.1-0.2 nM.

Claims: - 39 - What is claimed is: 1. An isolated polynucleotide comprising a nucleotide sequence that has at least 80% identity to a nucleotide sequence encoding the polypeptide of SEQ ID NO: 2 or the corresponding fragment thereof; or a nucleotide sequence complementary to said nucleotide sequence. 2. The polynucleotide of claim 1 which is DNA or RNA. 3. The polynucleotide of claim 1 wherein said nucleotide sequence is at least 80% identical to that contained in SEQ ID NO: l . 4. The polynucleotide of claim 3 wherein said nucleotide sequence is contained in SEQ ID NO:l. 5. The polynucleotide of claim 1 wherein said encoding nucleotide sequence encodes the polypeptide of SEQ ID NO: 2 or a fragment thereof. 6. A polynucleotide probe or primer comprising at least 15 contiguous nucleotides of the polynucleotide of claim 3. 7. A DNA or RNA molecule comprising an expression system wherein said expression system is capable of producing a Human GPR14 or a fragment thereof having at least 80% identity with a nucleotide sequence encoding the polypeptide of SEQ ID NO:2 or said fragment when said expression system is present in a compatible host ceU. 8. A host ceU comprising the expression system of claim 7. - 40 - 9. A process for producing a Human GPR14 polypeptide or fragment comprising culturing a host of claim 8 and under conditions sufficient for the production of said polypeptide or fragment. 10. The process of claim 9 wherein said polypeptide or fragment is expressed at the surface of said cell. 11. Cells produced by the process of claim 10. 12. The process of claim 9 which further includes recovering the polypeptide or fragment from the culture. 13. A process for producing a cell which produces a Human GPR14 polypeptide or a fragment thereof comprising transforming or transfecting a host cell with the expression system of claim 7 such that the host ceU, under appropriate culture conditions, produces a Human GPR14 polypeptide or fragment. 14. A Human GPR14 polypeptide or a fragment thereof comprising an amino acid sequence which is at least 85 % identical to the amino acid sequence contained in SEQ ID NO:2. 15. The polypeptide of claim 14 which comprises the amino acid sequence of SEQ ID NO:2, or a fragment thereof. 16. A Human GPR14 polypeptide or fragment prepared by the method of claim 12. - 41 - 17. An antibody immunospecific for the Human GPR14 polypeptide of claim 14. 18. A method for the treatment of a subject in need of enhanced Human GPR14 activity comprising: (a) administering to the subject a therapeuticaUy effective amount of an agonist to said receptor; and/or (b) providing to the subject Human GPR14 polynucleotide in a form so as to effect production of said receptor activity in vivo. 19. A method for the treatment of a subject having need to inhibit Human GPR 14 activity comprising: (a) administering to the subject a therapeuticaUy effective amount of an antagonist to said receptor; and/or (b) administering to the subject a nucleic acid molecule that inhibits the expression of the nucleotide sequence encoding said receptor; and/ or (c) administering to the subject a therapeuticaUy effective amount of a polypeptide that competes with said receptor for its ligand. 20. A process for diagnosing a disease or a susceptibiUty to a disease in a subject related to expression or activity of Human GPR14 in a subject comprising: (a) determining the presence or absence of a mutation in the nucleotide sequence encoding said Human GPR14 in the genome of said subject; and/or (b) analyzing for the presence or amount of the Human GPR 14 expression in a sample derived from said subject. - 42 - 21. A method for identifying compounds which bind to Human GPR14 comprising: (a) contacting cells of claim 11 with a candidate compound; and (b) assessing the abUity of said candidate compound to bind to said ceUs. 22. The method of claim 21 which further includes determining whether the candidate compound effects a signal generated by activation of the Human GPR14 polypeptide at the surface of the ceU, wherein a candidate compound which effects production of said signal is identified as an agonist. 23. A method for identifying agonist or antagonist of a Human GPR14 polypeptide which comprises: contacting a cell expressing on the surface thereof the polypeptide, said polypeptide being associated with a second component capable of providing a detectable signal in response to the binding of a compound to said polypeptide, with a compound to be screened under conditions to permit binding to the polypeptide; and determining whether the compound binds to and activates or inhibits the polypeptide by measuring the level of a signal generated from the interaction of the compound with the polypeptide. 24. A method of claim 23 which further comprises conducting the identification of agonist or antagonist in the presence of labeled or unlabeled, fish or human urotensin π. 25. A method for identifying agonist or antagonist of a Human GPR14 polypeptide which comprises: determining the inhibition of binding of a Ugand to ceUs which have the polypeptide on the surface thereof, or to ceU membranes containing the polypeptide, in - 43 - the presence of a candidate compound under conditions to permit binding to the polypeptide, and determining the amount of ligand bound to the polypeptide, such that a compound capable of causing reduction of binding of a ligand is an agonist or antagonist. 26. A method of claim 25 in which a Ugand is labeled or unlabeled, fish or human urotensin π . 27. A method for identifying agonist or antagonist of rat GPR14 receptor polypeptide which comprises: contacting a ceU expressing on the surface thereof the polypeptide, said polypeptide being associated with a second component capable of providing a detectable signal in response to the binding of a compound to said polypeptide, with a compound to be screened under conditions to permit binding to the polypeptide; and determining whether the compound binds to and activates or inhibits the polypeptide by measuring the level a signal generated from the interaction of the compound with the polypeptide. 28. A method of claim 27 which further comprises conducting the identification of agonist or antagonist in the presence of labeled or unlabeled, fish or human urotensin II . 29. A method for identifying agonist or antagonist of rat GPR14 receptor polypeptide which comprises: determining the inhibition of binding of a Ugand to cells which have the polypeptide on the surface thereof, or to ceU membranes containing the polypeptide, in the presence of a candidate compound under conditions to permit binding to the polypeptide, and determining the amount of Ugand bound to the polypeptide, such that a compound capable of causing reduction of binding of a Ugand is an agonist or antagonist. - 44 - 30. A method of claim 29 in which a Ugand is labeled or unlabeled, fish or human urotensin π. 31. An agonist identified by the method of claim 22. 32. The method of claim 21 which further includes contacting said cell with a known agonist for said Human GPR14; and determining whether the signal generated by said agonist is diminished in the presence of said candidate compound, wherein a candidate compound which effects a diminution in said signal is identified as an antagonist for said Human GPR14. 33. An antagonist identified by the method of claim 24.

Inventors: AIYAR NAMBI V, AL-BARAZANJI KAMAL, AMES ROBERT S, ARNOLD ANNE ROMANIC, BERGSMA DERK J, CHAMBERS JON, DOUGLAS STEPHEN A, FOLEY JAMES J, GOUT BERNARD, KHANDOUDI NASSIRAH, SARAU HENRY M, SHABON USMAN, WILLETTE ROBERT N, AIYAR, NAMBI, V., AL-BARAZANJI, KAMAL, AMES, ROBERT, S., ARNOLD, ANNE, ROMANIC, BERGSMA, DERK, J., CHAMBERS, JON, DOUGLAS, STEPHEN, A., FOLEY, JAMES, J., GOUT, BERNARD, KHANDOUDI, NASSIRAH, SARAU, HENRY, M., SHABON, USMAN, WILLETTE, ROBERT, N.
Applicants: SMITHKLINE BEECHAM CORP, SMITHKLINE BEECHAM LAB, SMITHKLINE BEECHAM PLC, SMITHKLINE BEECHAM CORPORATION, SMITHKLINE BEECHAM LABORATOIRES PHARMACEUTIQUES, SMITHKLINE BEECHAM P.L.C.
Application year: 1999
Main classification: C12N15
IPC codes: A61K31, A61K38, A61K48, A61P43, C07K14, C07K16, C12N1, C12N5, C12N15, C12P21, C12Q1, G01N33