Bak apoptotic pores involve a flexible C-terminal region and juxtaposition of the C-terminal transmembrane domains.
Bak and Bax mediate apoptotic cell death by oligomerizing and forming a pore in the mitochondrial outer membrane. Both proteins anchor to the outer membrane via a C-terminal transmembrane domain, although its topology within the apoptotic pore is not known. Cysteine-scanning mutagenesis and hydrophilic labeling confirmed that in healthy mitochondria the Bak α9 segment traverses the outer membrane, with 11 central residues shielded from labeling. After pore formation those residues remained shielded, indicating that α9 does not line a pore. Bak (and Bax) activation allowed linkage of α9 to neighboring α9 segments, identifying an α9:α9 interface in Bak (and Bax) oligomers. Although the linkage pattern along α9 indicated a preferred packing surface, there was no evidence of a dimerization motif. Rather, the interface was invoked in part by Bak conformation change and in part by BH3:groove dimerization. The α9:α9 interaction may constitute a secondary interface in Bak oligomers, as it could link BH3:groove dimers to high-order oligomers. Moreover, as high-order oligomers were generated when α9:α9 linkage in the membrane was combined with α6:α6 linkage on the membrane surface, the α6-α9 region in oligomerized Bak is flexible. These findings provide the first view of Bak carboxy terminus (C terminus) membrane topology within the apoptotic pore.
The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3050, Australia.|Department of Medical Biology, The University of Melbourne, Parkville, Victoria 3010, Australia.
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Iyer
The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3050, Australia.
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Bell
The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3050, Australia.|Department of Medical Biology, The University of Melbourne, Parkville, Victoria 3010, Australia.
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Westphal
The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3050, Australia.|Department of Medical Biology, The University of Melbourne, Parkville, Victoria 3010, Australia.
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Anwari
The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3050, Australia.|Department of Medical Biology, The University of Melbourne, Parkville, Victoria 3010, Australia.
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Gulbis
Department of Chemistry and Physics, La Trobe Institute for Molecular Sciences, La Trobe University, Melbourne, Victoria 3086, Australia.
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Smith
The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3050, Australia.|Department of Medical Biology, The University of Melbourne, Parkville, Victoria 3010, Australia.
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Dewson
The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3050, Australia.|Department of Medical Biology, The University of Melbourne, Parkville, Victoria 3010, Australia.
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Kluck
D020816:Amino Acid Motifs; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D006801:Humans; D051379:Mice; D051336:Mitochondrial Membranes / Q000378:metabolism*; D055503:Protein Multimerization; D050998:bcl-2 Homologous Antagonist-Killer Protein / Q000737:chemistry* / Q000378:metabolism*; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D050998:bcl-2 Homologous Antagonist-Killer Protein; D051028:bcl-2-Associated X Protein