Prokineticin polypeptides, related compositions and methods

Abstract

The invention provides isolated polypeptides that stimulate gastroeintestinal smooth muscle contraction, including human prokineticin 1 and human prokineticin 2 polypeptides, and functional fragments and modifications thereof. Also provided are methods of stimulating gastrointestinal smooth muscle contraction in a mammal, by administering to the mammal an effective amount of a prokineticin polypeptide. The invention also provides nucleic acid molecules encoding a prokineticin polypeptide, and antibodies that selectively bind a prokineticin polypeptide. Further provided are methods of identifying a prokineticin receptor ligand, agonist or antagonist.

1 - 52 . (canceled)  
     
     
         53 . An isolated prokineticin receptor antagonist, comprising an amino acid sequence at least 80% identical to amino acids 7 to 77 of SEQ ID NO:3, said sequence comprising; 
 (a) the 10 conserved cysteine residues of SEQ ID NO:3, and    (b) from 0 to 9 of C-terminal amino acids 78 to 86 of SEQ ID NO:3, wherein amino acids 1 to 6 of said antagonist do not consist of amino acids AVITGA (SEQ ID NO:21).    
     
     
         54 . The isolated prokineticin receptor antagonist of  claim 53 , comprising 6 or more amino acids N-terminal to the first conserved cysteine residue.  
     
     
         55 . The isolated prokineticin receptor antagonist of  claim 53 , comprising 7 or more amino acids N-terminal to the first conserved cysteine residue.  
     
     
         56 . The isolated prokineticin receptor antagonist of  claim 55 , wherein said 7 or more amino acids are MAVITGA (SEQ ID NO:20).  
     
     
         57 . The isolated prokineticin receptor antagonist of  claim 53 , comprising SEQ ID NO:18.  
     
     
         58 . The isolated prokineticin receptor antagonist of  claim 57 , consisting of SEQ ID NO:18.  
     
     
         59 . The isolated prokineticin receptor antagonist of  claim 53 , comprising 5 or fewer amino acids N-terminal to said first conserved cysteine residue.  
     
     
         60 . The isolated prokineticin receptor antagonist of  claim 59 , wherein said 5 or fewer amino acids consists of VITGA (SEQ ID NO:22).  
     
     
         61 . The isolated prokineticin receptor antagonist of  claim 53 , comprising SEQ ID NO:16.  
     
     
         62 . The isolated prokineticin receptor antagonist of  claim 61 , consisting of SEQ ID NO:16.  
     
     
         63 . The isolated prokineticin receptor antagonist of  claim 53 , wherein amino acid residues that differ from residues 7 to 77 of SEQ ID NO:3 are conservative substitutions thereof.  
     
     
         64 . The isolated prokineticin receptor antagonist of  claim 53 , wherein amino acid residues that differ from residues 7 to 77 of SEQ ID NO:3 consist of the corresponding residues from SEQ ID NO:6.  
     
     
         65 . The isolated prokineticin receptor antagonist of  claim 53 , comprising amino acids 7 to 77 of SEQ ID NO:3.  
     
     
         66 . The isolated prokineticin receptor antagonist of  claim 53 , comprising amino acids 7 to 77 of SEQ ID NO:13.  
     
     
         67 . The isolated prokineticin receptor antagonist of  claim 53 , further comprising a tag.  
     
     
         68 . The isolated prokineticin receptor antagonist of  claim 53 , which is detectably labeled.  
     
     
         69 . An isolated prokineticin receptor antagonist, comprising an amino acid sequence at least 80% identical to amino acids 7 to 77 of SEQ ID NO:6, said sequence comprising; 
 (a) the 10 conserved cysteine residues of SEQ ID NO:6, and    (b) from 0 to 4 of C-terminal amino acids 78 to 81 of SEQ ID NO:6,    wherein amino acids 1 to 6 of said antagonist do not consist of amino acids AVITGA (SEQ ID NO:21).    
     
     
         70 . The isolated prokineticin receptor antagonist of  claim 69 , comprising 6 or more amino acids N-terminal to the first conserved cysteine residue.  
     
     
         71 . The isolated prokineticin receptor antagonist of  claim 69 , comprising 7 or more amino acids N-terminal to the first conserved cysteine residue.  
     
     
         72 . The isolated prokineticin receptor antagonist of  claim 71 , wherein said 7 or more amino acids comprise MAVITGA (SEQ ID NO:20).  
     
     
         73 . The isolated prokineticin receptor antagonist of  claim 69 , comprising 5 or fewer amino acids N-terminal to the first conserved cysteine residue.  
     
     
         74 . The isolated prokineticin receptor antagonist of  claim 73 , wherein said 5 or fewer amino acids consists of VITGA (SEQ ID NO:22).  
     
     
         75 . The isolated prokineticin receptor antagonist of  claim 69 , wherein amino acid residues that differ from residues 7 to 77 of SEQ ID NO:6 are conservative substitutions thereof.  
     
     
         76 . The isolated prokineticin receptor antagonist of  claim 69 , wherein amino acid residues that differ from residues 7 to 77 of SEQ ID NO:6 consist of the corresponding residues from SEQ ID NO:3.  
     
     
         77 . The isolated prokineticin receptor antagonist of  claim 69 , comprising amino acids 7 to 77 of SEQ ID NO:6.  
     
     
         78 . The isolated prokineticin receptor antagonist of  claim 69 , comprising amino acids 7 to 77 of SEQ ID NO:14.  
     
     
         79 . The isolated prokineticin receptor antagonist of  claim 69 , further comprising a tag.  
     
     
         80 . The isolated prokineticin receptor antagonist of  claim 69 , which is detectably labeled.  
     
     
         81 . A pharmaceutical composition, comprising the isolated antagonist of  claim 53  and a pharmaceutically acceptable carrier.  
     
     
         82 . A pharmaceutical composition, comprising the isolated antagonist of  claim 69  and a pharmaceutically acceptable carrier.  
     
     
         83 . A nucleic acid molecule encoding the antagonist of  claim 53 .  
     
     
         84 . An expression vector comprising the nucleic acid molecule of  claim 83  operatively linked to a promoter of gene expression.  
     
     
         85 . A host cell comprising the expression vector of  claim 84 .  
     
     
         86 . A method of preparing the isolated antagonist of  claim 53 , comprising culturing the host cell of  claim 85  so as to express said antagonist, substantially purifying said antagonist, and refolding said antagonist.  
     
     
         87 . A nucleic acid molecule encoding the antagonist of  claim 69 .  
     
     
         88 . An expression vector comprising the nucleic acid molecule of  claim 87  operatively linked to a promoter of gene expression.  
     
     
         89 . A host cell comprising the expression vector of  claim 88 .  
     
     
         90 . A method of preparing the isolated antagonist of  claim 69 , comprising culturing the host cell of  claim 89  so as to express said antagonist, substantially purifying said antagonist, and refolding said antagonist.