Mutation to Bax beyond the BH3 Domain Disrupts Interactions with Pro-survival Proteins and Promotes Apoptosis
- Author(s)
- Czabotar, PE; Lee, EF; Thompson, GV; Wardak, AZ; Fairlie, WD; Colman, PM;
- Details
- Publication Year 2011-03-04,Volume 286,Issue #9,Page 7123-7131
- Journal Title
- JOURNAL OF BIOLOGICAL CHEMISTRY
- Publication Type
- Journal Article
- Abstract
- Pro-survival members of the Bcl-2 family of proteins restrain the pro-apoptotic activity of Bax, either directly through interactions with Bax or indirectly by sequestration of activator BH3-only proteins, or both. Mutations in Bax that promote apoptosis can provide insight into how Bax is regulated. Here, we describe crystal structures of the pro-survival proteins Mcl-1 and Bcl-x(L) in complex with a 34-mer peptide from Bax that encompasses its BH3 domain. These structures reveal canonical interactions between four signature hydrophobic amino acids from the BaxBH3 domain and the BH3-binding groove of the pro-survival proteins. In both structures, Met-74 from the Bax peptide engages with the BH3-binding groove in a fifth hydrophobic interaction. Various Bax Met-74 mutants disrupt interactions between Bax and all pro-survival proteins, but these Bax mutants retain pro-apoptotic activity. Bax/Bak-deficient mouse embryonic fibroblast cells reconstituted with several Bax Met-74 mutants are more sensitive to the BH3 mimetic compound ABT-737 as compared with cells expressing wild-type Bax. Furthermore, the cells expressing Bax Met-74 mutants are less viable in colony assays even in the absence of an external apoptotic stimulus. These results support a model in which direct restraint of Bax by pro-survival Bcl-2 proteins is a barrier to apoptosis.
- Publisher
- AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
- Keywords
- BCL-2 FAMILY-MEMBERS; CRYSTALLOGRAPHIC STRUCTURE DETERMINATION; PROGRAMMED CELL-DEATH; BH3-ONLY PROTEINS; MEMBRANE PERMEABILIZATION; X-RAY; CONFORMATIONAL-CHANGE; ANTIAPOPTOTIC BCL-2; ACTIVATION; BCL-X(L)
- Publisher's Version
- https://doi.org/10.1074/jbc.M110.161281
- Terms of Use/Rights Notice
- Refer to copyright notice on published article.
Creation Date: 2011-03-04 12:00:00