Membrane insertion of the BAX core, but not latch domain, drives apoptotic pore formation.

ABSTRACT: Despite intensive research effort, how the paradigmatic proapoptotic protein BAX forms lethal apoptotic pores at the mitochondrial outer membrane (MOM) remains incompletely understood. Here, we used biophysical tools and minimalist model systems to identify the specific regions in BAX driving apoptotic pore formation, and to gain more insight into underlying mechanisms. Fluorescence mapping revealed that fully active BAX adopts a BH3-in-groove dimeric conformation in MOM-like membranes, with BAX ?4-?5 helices belonging to its core domain inserting deeper into the membrane lipid bilayer than BAX ?6-?8 helices belonging to its latch domain. In our reconstituted systems, antiapoptotic BCLXL formed canonical heterodimeric BH3-in-groove complexes with BAX, and blocked membrane insertion of BAX core ?4-?5 helices, but not BAX latch ?6-?8 helices. Moreover, poly(ethylene glycol) (PEG) conjugation (PEGylation) at multiple individual sites along the BAX core, but not latch domain, potently inhibited BAX pore-forming activity. Additional combined computational and experimental evidence revealed that the BAX core ?5 helix displays a bilayer-destabilizing membrane interaction mode that is absent in BAX latch ?6-?8 helices. Based on this collective set of evidence, we propose that membrane insertion of the BAX core, but not latch domain, is critical for BAX apoptotic pore formation.

SUBMITTER: Flores-Romero H 

PROVIDER: S-EPMC5701199 | biostudies-literature | 2017 Nov

REPOSITORIES: biostudies-literature