Vesiculins

Abstract

The inventions relate generally to vesiculin peptides and vesiculin peptide chains, and fragments, variants and derivatives thereof, related compositions and formulations and their preparation and use, nucleic acids encoding such vesiculin peptides and vesiculin peptide chains, and fragments, variants and derivatives thereof and related vectors and host cells, hybridomas and antibodies, and methods for the prevention and treatment of conditions, diseases and disorders that would be improved, eased, or lessened by the administration of a composition of the invention, including but not limited to glucose metabolism diseases.

1 . An isolated polypeptide comprising a first chain having the amino acid sequence: 
       
         
           
                 
               
                   (SEQ ID NO: 3) 
                 
                   Gly Ile Val Glu Glu Cys 3  Cys 4  Phe Arg Ser Cys 6  Asp 
                 
                     
                 
                   Leu R 1  R 2  Leu Glu R 3  Tyr Cys 6  Ala R 4  R 5  R 6  R 7  R 8  R 9   
                 
             
                
                
                
                
               
            
           
         
       
       and a second chain having the amino acid sequence: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 58) 
                 
                     
                   R 10  R 11  R 12  R 13  R 14  Glu Thr Leu Cys 1  Gly Gly Glu Leu 
                 
                     
                     
                 
                     
                   Val Asp R 15  Leu Gln Phe R 16  Cys 2  R 17  R 18  Arg Gly 
                 
                     
                     
                 
                     
                   Phe Tyr Phe R 19  R 20  R 21  R 22  R 23  R 24  R 25  R 26  R 27  R 28  R 29   
                 
             
                
                
                
                
                
                
               
            
           
         
       
       wherein R 1  is Ala, Asn, Leu, or a conservative variant of either, R 2  is Leu, Ile, or a conservative variant of either, R 3  is Thr, Gln, or a conservative variant of either, R 4  is Thr, Ala, Lys, Val, or a conservative variant of either, R 5  is Pro, Ser, or a conservative variant of either, R 6  is Ala, Val, Pro, or a conservative variant thereof, R 7  is Lys, Glu, or a conservative variant of either, R 8  is Ser, Ala, or a conservative variant of either, R 9  is Glu, Ala, or a conservative variant of either, R 10  is absent or is Ala, Glu, Asp, or a conservative variant thereof, R 11  is Tyr, Ala, Val, or a conservative variant of either, R 12  is Arg, Gly, Ala, or a conservative variant thereof, R 13  is Pro, Thr, Ser, Leu, or a conservative variant thereof, R 14  is Ser, Gly, Ala, Glu, or a conservative variant thereof, R 15  is Thr, Tyr, Ala, or a conservative variant thereof, R 16  is Val, Ile, or a conservative variant thereof, R 17  is Gly, Ser, Glu, Ala, or a conservative variant thereof, R 18  is Asp, Glu, or a conservative variant of either, R 19  is Ser, Val, or a conservative variant of either, R 20  is Arg, Leu, Ser, or a conservative variant thereof, R 21  is Pro, Lys, or a conservative variant of either, R 22  is Ala, Ser, Gly, Val, Thr, or a conservative variant thereof, R 23  is Ser, Gly, Val, or a conservative variant thereof, R 24  is Arg, Pro, Gly, or a conservative variant thereof, R 25  is Ala, Arg, Val, Ile, Leu, Asn, Ser, Gly, or a conservative variant thereof, R 26  is Ser, Asn, Arg, or a conservative variant thereof, R 27  is absent or is Arg, Ser, Asn, or a conservative variant thereof, R 28  is absent or is Val, or a conservative variant thereof, R 29  is absent or is Ser, or a conservative variant thereof. 
     
     
         2 . The isolated polypeptide of  claim 1 , wherein the first and second chains are joined by one or more disulfide bonds. 
     
     
         3 . The isolated polypeptide of  claim 2 , wherein the first and second chains are joined by two disulfide bonds. 
     
     
         4 . A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a polypeptide according to  claim 3 . 
     
     
         5 . A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a polypeptide according to  claim 2 . 
     
     
         6 . A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a polypeptide according to  claim 1 . 
     
     
         7 . The polypeptide of  claim 1  wherein said polypeptide has 62 amino acids. 
     
     
         8 . The polypeptide of  claim 1  wherein said polypeptide has 63 amino acids. 
     
     
         9 . The polypeptide of  claim 1  wherein said polypeptide is at least about 90% pure. 
     
     
         10 . The polypeptide of  claim 1 , wherein said polypeptide is at least about 95% pure. 
     
     
         11 . The isolated polypeptide of  claim 1  comprising a first chain having the amino acid sequence: 
       
         
           
                 
               
                   (SEQ ID NO: 2) 
                 
                   Gly Ile Val Glu Glu Cys 3  Cys 4  Phe Arg Ser Cys 5  Asp 
                 
                     
                 
                   Leu Ala Leu Leu Glu Thr Tyr Cys 6  Ala R 7  Pro Ala 
                 
                     
                 
                   Lys Ser Glu 
                 
             
                
                
                
                
                
                
               
            
           
         
       
       and a second chain having the amino acid sequence: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 57) 
                 
                     
                   Ala 1  Tyr R 1  Pro R 2  Glu Thr Leu Cys 1  Gly Gly Glu 
                 
                     
                     
                 
                     
                   Leu Val Asp Tyr Leu Gln Phe Val Cys 2  R 3  Asp Arg 
                 
                     
                     
                 
                     
                   Gly Phe Tyr Phe Ser Arg Pro R 4  Ser Arg R 5  R 6   
                 
             
                
                
                
                
                
                
               
            
           
         
       
       wherein Ala 1  is either present or absent; R 1  is Gly or Arg, or a conservative variant of either; R 2  is Gly or Ser, or a conservative variant of either; R 3  is Gly or Ser, or a conservative variant of either; R 4  is Ser or Ala, or a conservative variant of either; R 5  is Ile or Val or Ala, or a conservative variant of either; R 6  is Asn or Ser, or a conservative variant of either; and R 7  is Thr or Ala, or a conservative variant of either. 
     
     
         12 . A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a polypeptide according to  claim 11 . 
     
     
         13 . The polypeptide of  claim 1  which is a human vesiculin. 
     
     
         14 . The polypeptide of  claim 13  comprising a first chain having the amino acid sequence: 
       
         
           
                 
               
                   (SEQ ID NO: 1) 
                 
                   Gly Ile Val Glu Glu Cys 3  Cys 4  Phe Arg Ser Cys 6  Asp 
                 
                     
                 
                   Leu Ala Leu Leu Glu Thr Tyr Cys 6  Ala Thr Pro Ala 
                 
                     
                 
                   Lys Ser Glu 
                 
             
                
                
                
                
                
                
               
            
           
         
       
       and a second chain having the amino acid sequence: 
       
         
           
                 
               
                   (SEQ ID NO: 56) 
                 
                   Ala 1  Tyr Arg Pro Ser Glu Thr Leu Cys 1  Gly Gly Glu 
                 
                     
                 
                   Leu Val Asp Thr Leu Gln Phe Val Cys 2  Gly Asp Arg 
                 
                     
                 
                   Gly Phe Tyr Phe Ser Arg Pro Ala Ser Arg Val Ser, 
                 
             
                
                
                
                
                
                
               
            
           
         
       
       wherein Ala 1  is either present or absent. 
     
     
         15 . A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a polypeptide according to  claim 14 . 
     
     
         16 . A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a polypeptide according to  claim 13 .