title: Evidence for an Induced-Fit Process Underlying the Activation of Apoptotic BAX by an Intrinsically Disordered BimBH3 Peptide.
abstract: Apoptotic BAX protein functions as a critical gateway to mitochondria-mediated apoptosis. A diversity of stimuli has been implicated in initiating BAX activation, but the triggering mechanism remains elusive. Here we study the interaction of BAX with an intrinsically disordered BH3 motif of Bim protein (BimBH3) using ESR techniques. Upon incubation with BAX, BimBH3 binds to BAX at helices 1/6 trigger site to initiate conformational changes of BAX, which in turn promotes the formation of BAX oligomers. The study strategy is twofold: while BAX oligomerization was monitored through spectral changes of spin-labeled BAX, the binding kinetics was studied by observing time-dependent changes of spin-labeled BimBH3. Meanwhile, conformational transition between the unstructured and structured BimBH3 was measured. We show that helical propensity of the BimBH3 is increased upon binding to BAX but is then reduced after being released from the activated BAX; the release is due to the BimBH3-induced conformational change of BAX that is a prerequisite for the oligomer assembling. Intermediate states are identified, offering a key snapshot of the coupled folding and binding process. Our results provide a quantitative mechanistic description of the BAX activation and reveal new insights into the mechanism underlying the interactions between BAX and BH3-mimetic peptide.
authors.affiliation: Department of Chemistry, National Tsing Hua University , Hsinchu 30013, Taiwan.
authors.forename: Siao-Ru
authors.initials: SR
authors.lastname: Jhong
authors.affiliation: Department of Chemistry, National Tsing Hua University , Hsinchu 30013, Taiwan.
authors.forename: Ching-Yu
authors.initials: CY
authors.lastname: Li
authors.affiliation: Department of Chemistry, National Tsing Hua University , Hsinchu 30013, Taiwan.
authors.forename: Tai-Ching
authors.initials: TC
authors.lastname: Sung
authors.affiliation: Department of Chemistry, National Tsing Hua University , Hsinchu 30013, Taiwan.
authors.forename: Yu-Jing
authors.initials: YJ
authors.lastname: Lan
authors.affiliation: Department of Chemistry, National Tsing Hua University , Hsinchu 30013, Taiwan.
authors.forename: Kuo-Jung
authors.initials: KJ
authors.lastname: Chang
authors.affiliation: Department of Chemistry, National Tsing Hua University , Hsinchu 30013, Taiwan.
authors.forename: Yun-Wei
authors.initials: YW
authors.lastname: Chiang
mesh_terms: D017209:Apoptosis; D000072224:Bcl-2-Like Protein 11 / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D051028:bcl-2-Associated X Protein / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000072224:Bcl-2-Like Protein 11; D010455:Peptides; D051028:bcl-2-Associated X Protein