Detergent-activated BAX protein is a monomer

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

  • Olena Ivashyna - , Washington University St. Louis (Autor:in)
  • Ana J. García-Sáez - , Technische Universität Dresden (Autor:in)
  • Jonas Ries - , Technische Universität Dresden (Autor:in)
  • Eric T. Christenson - (Autor:in)
  • Petra Schwille - , Technische Universität Dresden (Autor:in)
  • Paul H. Schlesinger - (Autor:in)

Abstract

BAX is a pro-apoptotic member of the BCL-2 protein family. At the onset of apoptosis, monomeric, cytoplasmic BAX is activated and translocates to the outer mitochondrial membrane, where it forms an oligomeric pore. The chemical mechanism of BAX activation is controversial, and several in vitro and in vivo methods of its activation are known. One of the most commonly used in vitro methods is activation with detergents, such as n-octyl glucoside. During BAX activation with n-octyl glucoside, it has been shown that BAX forms high molecular weight complexes that are larger than the combined molecular weight of BAX monomer and one detergent micelle. These large complexes have been ascribed to the oligomerization of BAX prior to its membrane insertion and pore formation. This is in contrast to the in vivo studies that suggest that active BAX inserts into the outer mitochondrial membrane as a monomer and then undergoes oligomerization. Here, to simultaneously determine the molecular weight and the number of BAX proteins per BAX-detergent micelle during detergent activation, we have used an approach that combines two single-molecule sensitivity technique, fluorescence correlation spectroscopy, and fluorescence-intensity distribution analysis. We have tested a range of detergents as follows: n-octyl glucoside, dodecyl maltoside, Triton X-100, Tween 20, 3-[(3-cholamidopropyl)dimethylammonio]-1-pro-panesulfonic acid, and cholic acid. With these detergents we observe that BAX is a monomer before, during, and after interaction with micelles. We conclude that detergent activation of BAX is not congruent with oligomerization and that in physiologic buffer conditions BAX can assume two stable monomeric conformations, one inactive and one active.

Details

OriginalspracheEnglisch
Seiten (von - bis)23935-23946
Seitenumfang12
FachzeitschriftJournal of Biological Chemistry
Jahrgang284
Ausgabenummer36
PublikationsstatusVeröffentlicht - 4 Sept. 2009
Peer-Review-StatusJa
Extern publiziertJa

Externe IDs

PubMed 19564333

Schlagworte