Assembly of the Bak apoptotic pore: a critical role for the Bak protein α6 helix in the multimerization of homodimers during apoptosis.
- Ma, S; Hockings, C; Anwari, K; Kratina, T; Fennell, S; Lazarou, M; Ryan, MT; Kluck, RM; Dewson, G;
Publication Year 2013-09-06, Volume 288, Issue #36, Page 26027-26038
- Journal Title
- The Journal of Biological Chemistry
- Publication Type
- Journal Article
- Bak and Bax are the essential effectors of the intrinsic pathway of apoptosis. Following an apoptotic stimulus, both undergo significant changes in conformation that facilitates their self-association to form pores in the mitochondrial outer membrane. However, the molecular structures of Bak and Bax oligomeric pores remain elusive. To characterize how Bak forms pores during apoptosis, we investigated its oligomerization under native conditions using blue native PAGE. We report that, in a healthy cell, inactive Bak is either monomeric or in a large complex involving VDAC2. Following an apoptotic stimulus, activated Bak forms BH3:groove homodimers that represent the basic stable oligomeric unit. These dimers multimerize to higher-order oligomers via a labile interface independent of both the BH3 domain and groove. Linkage of the α6:α6 interface is sufficient to stabilize higher-order Bak oligomers on native PAGE, suggesting an important role in the Bak oligomeric pore. Mutagenesis of the α6 helix disrupted apoptotic function because a chimera of Bak with the α6 derived from Bcl-2 could be activated by truncated Bid (tBid) and could form BH3:groove homodimers but could not form high molecular weight oligomers or mediate cell death. An α6 peptide could block Bak function but did so upstream of dimerization, potentially implicating α6 as a site for activation by BH3-only proteins. Our examination of native Bak oligomers indicates that the Bak apoptotic pore forms by the multimerization of BH3:groove homodimers and reveals that Bak α6 is not only important for Bak oligomerization and function but may also be involved in how Bak is activated by BH3-only proteins.
- Apoptosis ; Bax ; Bcl-2 Family Proteins ; Mitochondria ; Protein Conformation ; Bak ; Blue Native PAGE ; Oligomerization
- WEHI Research Division(s)
- Molecular Genetics Of Cancer; Cell Signalling And Cell Death
- Link To PubMed Central Version
- Publisher's Version
- Rights Notice
- © 2013 by The American Society for Biochemistry and Molecular Biology
Creation Date: 2013-09-06 12:00:00Last Modified: 0001-01-01 12:00:00