title: Oligomerization process of Bcl-2 associated X protein revealed from intermediate structures in solution.
abstract: Upon apoptotic stress, Bcl-2 associated X (BAX) protein undergoes conformational changes and oligomerizes, leading to the mitochondrial membrane permeabilization and cell death. While structures of the resultant oligomer have been extensively studied, little is known about the intermediates that describe the reaction pathway from the inactive monomers to activated oligomers. Here we characterize the intermediate structures of BAX using combined small-angle X-ray scattering (SAXS) with on-line gel-filtration and electron spin resonance (ESR). The intermediates, including monomers, dimers, and tetramers, are reconstructed via integrating the SAXS-envelopes and ESR-determined skeleton structures. The hence revealed structures suggest a linear oligomerization of BAX utilizing the extended dimers with the two flexible α6 chains protruded out as ditopic ligands. The results of molecular dynamics simulation also support the ditopic dimer conformation with mobile α6. The ditopic dimers could further wind into a helical rod structure with three dimers in one helical turn. Our results not only reveal the on-pathway intermediates, but also suggest a ditopic oligomerization mechanism that may bridge the observed intermediate structures in solution to the large BAX assemblies lately observed on mitochondria.
authors.affiliation: National Synchrotron Radiation Research Center, Hsinchu 30076, Taiwan. usjeng@nsrrc.org.tw.
authors.forename: Orion
authors.identifier: http://orcid.org/0000-0002-2247-5061
authors.initials: O
authors.lastname: Shih
authors.affiliation: National Synchrotron Radiation Research Center, Hsinchu 30076, Taiwan. usjeng@nsrrc.org.tw.
authors.forename: Yi-Qi
authors.initials: YQ
authors.lastname: Yeh
authors.affiliation: National Synchrotron Radiation Research Center, Hsinchu 30076, Taiwan. usjeng@nsrrc.org.tw.
authors.forename: Kuei-Fen
authors.initials: KF
authors.lastname: Liao
authors.affiliation: Department of Chemistry, National Tsing Hua University, Hsinchu 30013, Taiwan. ywchiang@gapp.nthu.edu.tw.
authors.forename: Tai-Ching
authors.initials: TC
authors.lastname: Sung
authors.affiliation: Department of Chemistry, National Tsing Hua University, Hsinchu 30013, Taiwan. ywchiang@gapp.nthu.edu.tw.
authors.forename: Yun-Wei
authors.identifier: http://orcid.org/0000-0002-2101-8918
authors.initials: YW
authors.lastname: Chiang
authors.affiliation: National Synchrotron Radiation Research Center, Hsinchu 30076, Taiwan. usjeng@nsrrc.org.tw and Department of Chemical Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan.
authors.forename: U-Ser
authors.initials: US
authors.lastname: Jeng
mesh_terms: D000595:Amino Acid Sequence; D004578:Electron Spin Resonance Spectroscopy; D006801:Humans; D056004:Molecular Dynamics Simulation; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D053838:Scattering, Small Angle; D012996:Solutions / Q000737:chemistry*; D013053:Spectrophotometry; D014961:X-Ray Diffraction; D051028:bcl-2-Associated X Protein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D012996:Solutions; D051028:bcl-2-Associated X Protein