Methods for identifying modulators of apoptosis
First Claim
1. A method of identifying an effective compound that decreases the binding of Humanin to Bax, comprising the steps of:
- (a) contacting said Humanin, wherein Humanin comprises the amino acid sequence SEQ ID NO;
2 or SEQ ID NO;
3, with said Bax, wherein Bax comprises the amino acid sequence SEQ ID NO;
5, in vitro under conditions suitable to form a Humanin-Bax complex;
(b) contacting said Humanin-Bax complex with a candidate compound; and
(c) determining the ability of said candidate compound to decrease the binding of said Humanin to said Bax, wherein decreasing the binding of said Humanin to said Bax indicates that said candidate compound is an effective compound that decreases the binding of said Humanin to said Bax.
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Abstract
The invention provides a method of identifying an effective compound that modulates the binding of Humanin to Bax or Bid. The invention also provides a method of identifying an effective compound that modulates an activity of Bax or Bid. In addition, the invention provides a method of identifying a Humanin-like compound that binds to Bax or Bid or modulates an activity of Bax or Bid, or inhibits the apoptotic activity of Bax or Bid. The invention further provides an isolated polypeptide containing a mitochondrial-derived form of Humanin (SEQ ID NO:3) or a functional fragment thereof where the fragment contains the methionine at position 16 of SEQ ID NO:3.
28 Citations
13 Claims
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1. A method of identifying an effective compound that decreases the binding of Humanin to Bax, comprising the steps of:
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(a) contacting said Humanin, wherein Humanin comprises the amino acid sequence SEQ ID NO;
2 or SEQ ID NO;
3, with said Bax, wherein Bax comprises the amino acid sequence SEQ ID NO;
5, in vitro under conditions suitable to form a Humanin-Bax complex;(b) contacting said Humanin-Bax complex with a candidate compound; and (c) determining the ability of said candidate compound to decrease the binding of said Humanin to said Bax, wherein decreasing the binding of said Humanin to said Bax indicates that said candidate compound is an effective compound that decreases the binding of said Humanin to said Bax. - View Dependent Claims (2, 3, 4, 5, 10, 11)
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6. A method of identifying an effective compound that decreases the binding of Humanin to Bax, comprising the steps of:
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(a) contacting said Humanin, wherein Humanin comprises the amino acid sequence SEQ ID NO;
2 or SEQ ID NO;
3, with said Bax, wherein Bax comprises the amino acid sequence SEQ ID NO;
5, under conditions suitable to form a Humanin-Bax complex in a yeast two-hybrid assay;(b) contacting said Humanin-Bax complex with a candidate compound; and (c) determining the ability of said candidate compound to decrease the binding of said Humanin to said Bax, wherein decreasing the binding of said Humanin to said Bax indicates that said candidate compound is an effective compound that decreases the binding of said Humanin to said Bax. - View Dependent Claims (7, 8, 9, 12, 13)
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Specification