title: Three-dimensional structure of Bax-mediated pores in membrane bilayers.
abstract: B-cell lymphoma 2 (Bcl-2)-associated X protein (Bax) is a member of the Bcl-2 protein family having a pivotal role in triggering cell commitment to apoptosis. Bax is latent and monomeric in the cytosol but transforms into its lethal, mitochondria-embedded oligomeric form in response to cell stress, leading to the release of apoptogenic factors such as cytochrome C. Here, we dissected the structural correlates of Bax membrane insertion while oligomerization is halted. This strategy was enabled through the use of nanometer-scale phospholipid bilayer islands (nanodiscs) the size of which restricts the reconstituted system to single Bax-molecule activity. Using this minimal reconstituted system, we captured structural correlates that precede Bax homo-oligomerization elucidating previously inaccessible steps of the core molecular mechanism by which Bcl-2 family proteins regulate membrane permeabilization. We observe that, in the presence of BH3 interacting domain death agonist (Bid) BH3 peptide, Bax monomers induce the formation of ~3.5-nm diameter pores and significantly distort the phospholipid bilayer. These pores are compatible with promoting release of ions as well as proteinaceous components, suggesting that membrane-integrated Bax monomers in the presence of Bid BH3 peptides are key functional units for the activation of the cell demolition machinery.
authors.affiliation: Bioinformatics and Systems Biology Program, Sanford-Burnham Medical Research Institute, La Jolla, CA 92037, USA.
authors.forename: X-P
authors.initials: XP
authors.lastname: Xu
authors.forename: D
authors.initials: D
authors.lastname: Zhai
authors.forename: E
authors.initials: E
authors.lastname: Kim
authors.forename: M
authors.initials: M
authors.lastname: Swift
authors.forename: J C
authors.initials: JC
authors.lastname: Reed
authors.forename: N
authors.initials: N
authors.lastname: Volkmann
authors.forename: D
authors.initials: D
authors.lastname: Hanein
mesh_terms: D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry; D051023:BH3 Interacting Domain Death Agonist Protein / Q000737:chemistry; D002463:Cell Membrane Permeability; D020285:Cryoelectron Microscopy; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry*; D049329:Nanostructures / Q000737:chemistry / Q000648:ultrastructure; D010446:Peptide Fragments / Q000737:chemistry; D016062:Porosity; D020836:Protein Structure, Quaternary; D051028:bcl-2-Associated X Protein / Q000737:chemistry* / Q000648:ultrastructure
chemical_list: C494826:BAX protein, human; D051023:BH3 Interacting Domain Death Agonist Protein; C494814:BID protein, human; D008051:Lipid Bilayers; D010446:Peptide Fragments; D051028:bcl-2-Associated X Protein; D015060:1,2-Dipalmitoylphosphatidylcholine