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US 6,607,880 B1Granted
Methods and reagents for modulating apoptosis
AssigneeMcGill University
InventorsGordon C. Shore, Florence W. H. Ng, Mai Nguyen, Philip E. Branton
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Abstract
The invention provides a novel apoptosis protein: p28 Bap31. Also provided are p28 Bap31 polypeptide fragments, p28 Bap31 antisense nucleic acid molecules, anti-p28 Bap31 antibodies, and methods for modulating apoptosis and for detecting compounds which modulate apoptosis.
Claims21
A method for identifying a compound that modulates apoptosis, said method comprising the steps of:
- (a) providing a cell having:
- (i) a reporter gene operably linked to a DNA-binding-protein recognition site;
- (ii) a first fusion gene capable of expressing a first fusion protein comprising a polypeptide covalently bonded to a polypeptide which is a binding moiety that is capable of specifically binding to said DNA-binding-protein recognition site;
- and
- (iii) a second fusion gene capable of expressing a second fusion protein comprising a polypeptide of a second protein covalently bonded to a gene activating moiety, wherein either said first polypeptide or said second polypeptide is human p28 Bap31 (SEQ ID NO:
- 1) or mouse p28 Bap31 (SEQ ID NO:
- 33) or a fragment of said human or mouse p28 Bap31, said fragment comprising a domain required for association of p28 Bap31 with pro-FLICE or a domain required for association of p28 Bap31 with Bcl-2 or Bcl-XL;
- (b) exposing said cell to said compound;
- and
- (c) measuring reporter gene expression in said cell, a change in said reporter gene expression identifying said candidate compound as a compound that modulates apoptosis.
7 more elements
A method for identifying a compound that modulates apoptosis, said method comprising:
- (a) providing human p28 Bap31 (SEQ ID NO:
- 1) or mouse p28 Bap31 (SEQ ID NO:
- 33) or a fragment of said human or mouse p28 Bap31, said fragment comprising a domain required for association of p28 Bap31 with pro-FLICE or a domain required for association of p28 Bap31 with Bcl-2 or Bcl-XL;
- and
- (b) contacting said polypeptide with a candidate compound and monitoring the level of p28 Bap31 biological activity, wherein a change in said level of p28 Bap31 biological activity in response to said candidate compound relative to the level of p28 Bap31 biological activity of said p28 Bap31 polypeptide not contacted with said candidate compound identifies said candidate compound as a compound that modulates apoptosis.
2 more elements
A method for identifying a compound that modulates apoptosis, said method comprising the steps of:
- (a) contacting a first polypeptide comprising human p28 Bap31 (SEQ ID NO:
- 1) or mouse p28 Bap31 (SEQ ID NO:
- 33) or a fragment of said human or mouse p28 Bap31, said fragment comprising a domain required for association of p28 Bap31 with pro-FLICE or a domain required for association of p28 Bap31 with Bcl-2 or Bcl-XL with a second polypeptide that interacts with said p28 Bap31 polypeptide such that said first polypeptide and said second polypeptide can form an interaction;
- (c) contacting said first polypeptide and said second polypeptide with a candidate compound;
- and
- (d) measuring said interaction of said first polypeptide and said second polypeptide, a change in said interaction of said first polypeptide and said second polypeptide in the presence of said candidate compound relative to an interaction of said first polypeptide and said second polypeptide not contacted with said candidate compound identifying said compound as a compound that modulates apoptosis.
3 more elements
Dependencies are inferred from the claim text.