Bax forms an oligomer via separate, yet interdependent, surfaces.
Interactions of Bcl-2 family proteins regulate permeability of the mitochondrial outer membrane and apoptosis. In particular, Bax forms an oligomer that permeabilizes the membrane. To map the interface of the Bax oligomer we used Triton X-100 as a membrane surrogate and performed site-specific photocross-linking. Bax-specific adducts were formed through photo-reactive probes at multiple sites that can be grouped into two surfaces. The first surface overlaps with the BH1-3 groove formed by Bcl-2 Homology motif 1, 2, and 3; the second surface is a rear pocket located on the opposite side of the protein from the BH1-3 groove. Further cross-linking experiments using Bax BH3 peptides and mutants demonstrated that the two surfaces interact with their counterparts in neighboring proteins to form two separated interfaces and that interaction at the BH1-3 groove primes the rear pocket for further interaction. Therefore, Bax oligomerization proceeds through a series of interactions that occur at separate, yet allosterically, coupled interfaces.
Zhang
Zhi
Z
Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73126, USA.
Zhu
Weijia
W
Lapolla
Suzanne M
SM
Miao
Yiwei
Y
Shao
Yuanlong
Y
Falcone
Mina
M
Boreham
Doug
D
McFarlane
Nicole
N
Ding
Jingzhen
J
Johnson
Arthur E
AE
Zhang
Xuejun C
XC
Andrews
David W
DW
Lin
Jialing
J
D000495:Allosteric Site; D020816:Amino Acid Motifs; D017209:Apoptosis; D001671:Biochemistry / Q000379:methods; D003432:Cross-Linking Reagents / Q000737:chemistry; D003902:Detergents / Q000494:pharmacology; D006801:Humans; D009154:Mutation; D017830:Octoxynol / Q000494:pharmacology; D010455:Peptides / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000737:chemistry; D051028:bcl-2-Associated X Protein / Q000378:metabolism*
D003432:Cross-Linking Reagents; D003902:Detergents; D010455:Peptides; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D017830:Octoxynol
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