Neuropeptide Y receptor Y5 and nucleic acid sequences

Abstract

The present invention provides novel NPY/PYY receptor proteins and the nucleic acid sequence encoding them. The invention is directed to the isolation, characterization, and pharmacological use of these receptors and nucleic acids. In particular, this invention provides human and rat NPY/PYY receptors (which we call the NPY Y5 receptor) and nucleic acids. Also provided are recombinant expression constructs useful for transfecting cells and expressing the protein in vitro and in vivo. The invention further provides methods for detecting expression levels of the protein as well as methods for screening for receptor antagonists and agonists to be used for the treatment of obesity or anorexia, respectively.

We claim:  
     
         1 . An isolated nucleic acid encoding a neuropeptide Y receptor comprising a molecule having a nucleotide sequence substantially the same as SEQ ID NO 1, SEQ ID NO 3, or SEQ ID NO 5.  
     
     
         2 . A homogenous composition of the neuropeptide Y receptor comprising a molecule having an amino acid sequence substantially the same as SEQ ID NO 2, SEQ ID NO 4, or SEQ ID NO 6.  
     
     
         3 . A vector comprising the nucleic acid according to  claim 1 .  
     
     
         4 . A vector according to  claim 3  adapted for expression in a cell further comprising regulatory elements necessary for expression of the nucleic acid operatively linked to the nucleic acid to enable expression of the nucleic acid.  
     
     
         5 . A vector according to  claim 4  wherein the cell is a mammalian cell.  
     
     
         6 . A vector according to  claim 5  wherein the cell is a human 293 cell.  
     
     
         7 . A vector according to  claim 4  that is a plasmid.  
     
     
         8 . A vector according to  claim 7  wherein the plasmid is the pBluescript plasmid.  
     
     
         9 . A vector according to  claim 7  wherein the plasmid is the pcDNA3 plasmid.  
     
     
         10 . A vector according to  claim 3  which is self-replicating.  
     
     
         11 . A cell transformed with the nucleic acid according to  claim 1  that expresses the nucleic acid.  
     
     
         12 . A cell according to  claim 11  that is a bacterium cell, an insect cell, or a yeast cell.  
     
     
         13 . A cell according to  claim 11  that is a mammalian cell.  
     
     
         14 . A cell according to  claim 13  that is a human 293 cell.  
     
     
         15 . A nucleic acid probe comprising a nucleic acid complementary to the nucleic acid according to  claim 1 .  
     
     
         16 . An antisense oligonucleotide having a sequence complementary to the nucleic acid according to  claim 1  and that inhibits expression of the nucleic acid.  
     
     
         17 . A membrane or membrane preparation comprising a membrane or portion thereof of a cell expressing a nucleic acid having a nucleotide sequence substantially the same as SEQ ID NO 1, SEQ ID NO 3, or SEQ ID NO 5.  
     
     
         18 . A membrane or membrane preparation according to  claim 17  wherein the cell is a mammalian cell.  
     
     
         19 . An antibody or fragment thereof that is immunologically reactive to a mammalian Y5 receptor.  
     
     
         20 . An antibody or fragment according to  claim 19  thereof wherein the mammalian Y5 receptor has an amino acid sequence substantially the same as one chosen from the group consisting of SEQ ID NO 2, SEQ ID NO 4, and SEQ ID NO 6.  
     
     
         21 . An antibody or fragment thereof according to  claim 19  that is a monoclonal antibody.  
     
     
         22 . An antibody or fragment thereof according to  claim 20  that is a monoclonal antibody.  
     
     
         23 . A cell line producing an antibody according to  claim 19 .  
     
     
         24 . A cell line producing an antibody according to  claim 20 .  
     
     
         25 . A cell line producing an antibody according to  claim 21 .  
     
     
         26 . A cell line producing an antibody according to  claim 22 .  
     
     
         27 . An epitope of a mammalian Y5 receptor wherein the epitope is immunologically reactive to the antibody or fragment thereof according to  claim 19 .  
     
     
         28 . An epitope of a mammalian Y5 receptor wherein the epitope is immunologically reactive to the antibody or fragment thereof according to  claim 20 .  
     
     
         29 . An epitope of a mammalian Y5 receptor wherein the epitope is immunologically reactive to the antibody or fragment thereof according to  claim 21 .  
     
     
         30 . An epitope of a mammalian Y5 receptor wherein the epitope is immunologically reactive to the antibody or fragment thereof according to  claim 22 .  
     
     
         31 . A method of producing a neuropeptide Y receptor comprising culturing a cell according to  claim 11  and recovering the receptor expressed by the cell.  
     
     
         32 . A method of producing a neuropeptide Y receptor comprising culturing a cell according to  claim 12  and recovering the receptor expressed by the cell.  
     
     
         33 . A method of producing a neuropeptide Y receptor comprising culturing a cell according to  claim 13  and recovering the receptor expressed by the cell.  
     
     
         34 . A method of producing a neuropeptide Y receptor comprising culturing a cell according to  claim 14  and recovering the receptor expressed by the cell.  
     
     
         35 . A method of identifying a neuropeptide Y agonist or antagonist comprising contacting a potential agonist or antagonist molecule with a membrane or membrane preparation according to  claim 17 .  
     
     
         36 . A method of identifying a neuropeptide Y agonist or antagonist comprising contacting a potential agonist or antagonist molecule with a membrane or membrane preparation according to  claim 18 .  
     
     
         37 . A neuropeptide Y antagonist comprising a compound identified according to  claim 36 .  
     
     
         38 . A method of suppressing the appetite of a mammal comprising administering to the mammal an appetite suppressing amount of a neuropeptide Y antagonist according to  claim 37 .  
     
     
         39 . A method of suppressing the appetite of a mammal according to  claim 38  wherein the amount of antagonist is from about 0.01 to about 100 mg/kg.  
     
     
         40 . A pharmaceutical composition comprising an effective appetite suppressing amount of an antagonist according to  claim 37  together with a pharmaceutically acceptable carrier.  
     
     
         41 . A neuropeptide Y agonist comprising a compound identified according to  claim 36 .  
     
     
         42 . A method of stimulating the appetite of a mammal comprising administering to the mammal an appetite stimulating amount of a neuropeptide Y agonist according to  claim 41 .  
     
     
         43 . A method of stimulating the appetite of a mammal according to  claim 42  wherein the amount of agonist is from about 0.01 to about 100 mg/kg.  
     
     
         44 . A pharmaceutical composition comprising an effective appetite stimulating amount of an agonist according to  claim 41  together with a pharmaceutically acceptable carrier.  
     
     
         45 . A non-human transgenic mammal that expresses the nucleic acid having a sequence substantially the same one chosen from the group consisting of SEQ ID NO 1, SEQ ID NO 3, and SEQ ID NO 5.