title: Multiple partners can kiss-and-run: Bax transfers between multiple membranes and permeabilizes those primed by tBid.
abstract: During apoptosis Bid and Bax are sufficient for mitochondrial outer membrane permeabilization, releasing pro-apoptotic proteins such as cytochrome c and Smac/Diablo into the cytoplasm. In most cells, both Bid and Bax are cytoplasmic but bind to mitochondrial outer membranes to exert pro-apoptotic functions. Binding to membranes is regulated by cleavage of Bid to truncated Bid (tBid), by conformation changes in tBid and Bax, and by interactions with other proteins. At least at the peripherally bound stage, binding is reversible. Therefore, regulation of apoptosis is closely linked with the interactions of tBid and Bax with mitochondria. Here we use fluorescence techniques and cell-free systems containing mitochondria or liposomes that faithfully mimic tBid/Bax-dependent membrane permeabilization to study the dynamic interactions of the proteins with membranes. We confirm that the binding of both proteins to the membrane is reversible by quantifying the binding affinity of proteins for the membrane. For Bax, both peripherally bound (inactive) and oligomerized (active) proteins migrate between membranes but much slower than and independent of tBid. When re-localized to a new membrane, Bax inserts into and permeabilizes it only if primed by an activator. In the case of tBid, the process of transfer is synergetic with Bax in the sense that tBid 'runs' faster if it has been 'kissed' by Bax. Furthermore, Mtch2 accelerates the re-localization of tBid at the mitochondria. In contrast, binding to Bcl-XL dramatically impedes tBid re-localization by lowering the off-rate threefold. Our results suggest that the transfer of activated tBid and Bax to different mitochondria is governed by dynamic equilibria and potentially contributes more than previously anticipated to the dissemination of the permeabilization signal within the cell.
authors.affiliation: Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada.
authors.forename: A
authors.initials: A
authors.lastname: Shamas-Din
authors.affiliation: Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada.
authors.forename: D
authors.initials: D
authors.lastname: Satsoura
authors.affiliation: Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada.
authors.forename: O
authors.initials: O
authors.lastname: Khan
authors.affiliation: Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada.
authors.forename: W
authors.initials: W
authors.lastname: Zhu
authors.affiliation: 1] Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada [2] Department of Medicine, McMaster University, Hamilton, ON, Canada.
authors.forename: B
authors.initials: B
authors.lastname: Leber
authors.affiliation: 1] Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada [2] Department of Physics and Astronomy, McMaster University, Hamilton, ON, Canada.
authors.forename: C
authors.initials: C
authors.lastname: Fradin
authors.affiliation: 1] Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada [2] Sunnybrook Research Institute and Department of Biochemistry, University of Toronto, Toronto, ON, Canada.
authors.forename: D W
authors.initials: DW
authors.lastname: Andrews
mesh_terms: D000818:Animals; D051023:BH3 Interacting Domain Death Agonist Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D051336:Mitochondrial Membranes / Q000737:chemistry / Q000378:metabolism*; D010539:Permeability; D051028:bcl-2-Associated X Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051020:bcl-X Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: C494826:BAX protein, human; C494811:BCL2L1 protein, human; D051023:BH3 Interacting Domain Death Agonist Protein; C494814:BID protein, human; C494827:Bax protein, mouse; C494812:Bcl2l1 protein, mouse; C494815:Bid protein, mouse; D008081:Liposomes; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein