Bax-mediated apoptosis modulating reagents and methods

Abstract

The invention features substantially purified polypeptide fragments of two specific domains, namely the ART domain and the transmembrane domain, within the BAX protein which, when administered to a cell, increase or decrease apoptosis of the cell. Also disclosed are methods for identifying compounds which, when administered to a cell, increase or decrease apoptosis of the cell. These compounds are identified based on their abilities to interact with specific domains of the BAX protein, or to alter the interaction of the BAX ART domain with the BAX transmembrane domain. In addition, the invention provides methods for diagnosing a patient having, or predisposed to develop, a disease involving altered apoptosis by identifying a mutation in a BAX-encoding gene which results in an amino acid mutation in the BAX ART domain, a BAX transmembrane domain, or that alters the interaction of the BAX ART domain with the BAX transmembrane domain.

What is claimed is:  
     
         1 . A substantially pure polypeptide consisting essentially of a BAX ART domain.  
     
     
         2 . The polypeptide of  claim 1 , wherein said BAX is human BAX, mouse BAX, or rat BAX.  
     
     
         3 . The polypeptide of  claim 1 , wherein said polypeptide decreases apoptosis of a cell when administered to said cell.  
     
     
         4 . The polypeptide of  claim 3 , wherein said cell is a degenerative cell.  
     
     
         5 . The polypeptide of  claim 3 , wherein said cell is a human cell.  
     
     
         6 . The polypeptide of  claim 1 , wherein said polypeptide has the sequence MDGSG(E/D)(Q/H)(L/P)(R/G)(S/G)GGPTSSEQI (SEQ ID NO: 1).  
     
     
         7 . A recombinant nucleic acid molecule encoding a polypeptide consisting essentially of a BAX ART domain.  
     
     
         8 . The nucleic acid molecule of  claim 7 , wherein said BAX is human BAX, mouse BAX, or rat BAX.  
     
     
         9 . The nucleic acid molecule of  claim 7 , wherein said polypeptide has the sequence MDGSG(E/D)(Q/H)(L/P)(R/G)(S/G)GGPTSSEQI (SEQ ID NO: 1).  
     
     
         10 . A pharmaceutical composition comprising a substantially pure polypeptide consisting essentially of a BAX ART domain and a pharmaceutically acceptable excipient.  
     
     
         11 . The pharmaceutical composition of  claim 10 , wherein said polypeptide decreases apoptosis of a cell when administered to said cell.  
     
     
         12 . The pharmaceutical composition of  claim 11 , wherein said cell is a degenerative cell.  
     
     
         13 . The pharmaceutical composition of  claim 11 , wherein said cell is a human cell.  
     
     
         14 . The pharmaceutical composition of  claim 10 , wherein said BAX is human BAX, mouse BAX, or rat BAX.  
     
     
         15 . The pharmaceutical composition of  claim 10 , wherein said polypeptide has the sequence MDGSG(E/D)(Q/H)(L/P)(R/G)(S/G)GGPTSSEQI (SEQ ID NO: 1).  
     
     
         16 . A method for decreasing apoptosis of a cell, said method comprising the step of contacting said cell with a substantially pure polypeptide consisting essentially of a BAX ART domain.  
     
     
         17 . The method of  claim 16 , wherein said cell is a degenerative cell.  
     
     
         18 . The method of  claim 16 , wherein said cell is a human cell.  
     
     
         19 . The method of  claim 16 , wherein said BAX is human BAX, mouse BAX, or rat BAX.  
     
     
         20 . The method of  claim 16 , wherein said polypeptide has the sequence MDGSG(E/D)(Q/H)(L/P)(R/G)(S/G)GGPTSSEQI (SEQ ID NO: 1).  
     
     
         21 . A method for decreasing apoptosis of a cell, said method comprising the step of expressing in said cell a recombinant nucleic acid molecule encoding a polypeptide consisting essentially of a BAX ART domain.  
     
     
         22 . The method of  claim 21 , wherein said cell is a degenerative cell.  
     
     
         23 . The method of  claim 21 , wherein said cell is a human cell.  
     
     
         24 . The method of  claim 21 , wherein said BAX is human BAX, mouse BAX, or rat BAX.  
     
     
         25 . The method of  claim 21 , wherein said polypeptide has the sequence MDGSG(E/D)(Q/H)(L/P)(R/G)(S/G)GGPTSSEQI (SEQ ID NO: 1).  
     
     
         26 . A method for decreasing apoptosis in a mammal, said method comprising administering to said mammal a substantially pure polypeptide consisting essentially of a BAX ART domain.  
     
     
         27 . The method of  claim 26 , wherein said mammal is a human.  
     
     
         28 . The method of  claim 26 , wherein said BAX is human BAX, mouse BAX, or rat BAX.  
     
     
         29 . The method of  claim 26 , wherein said polypeptide has the sequence MDGSG(E/D)(Q/H)(L/P)(R/G)(S/G)GGPTSSEQI (SEQ ID NO: 1).