Oligomerization of membrane-bound Bcl-2 is involved in its pore formation induced by tBid

Apoptosis. 2009 Oct;14(10):1145-53. doi: 10.1007/s10495-009-0389-8.

Abstract

Both pro-apoptotic Bax and anti-apoptotic Bcl-2 are structurally homologous to the pore-forming domain of bacterial toxins. Bax proteins oligomerize in the mitochondrial outer membranes forming pores that release cytochrome c from the mitochondrial intermembrane space. Bcl-2 proteins also form pores that, however, are much smaller than the Bax pore. It is unknown whether Bcl-2 forms monomeric or oligomeric pores. Here, we characterized the Bcl-2 pore formation in liposomes using biophysical and biochemical techniques. The results show that the Bcl-2 pore enlarges as the concentration of Bcl-2 increases, suggesting that the pore is formed by Bcl-2 oligomers. As expected from oligomerization-mediated pore-formation, the small pores are formed earlier than the large ones. Bcl-2 oligomers form pores faster than the monomer, indicating that the oligomerization constitutes an intermediate step of the pore formation. A Bcl-2 mutant with higher affinity for oligomerization forms pores faster than wild type Bcl-2. Bcl-2 oligomers were detected in the liposomal membranes under conditions that Bcl-2 forms pores, and the extent of oligomerization was positively correlated with the pore-forming activity. Therefore, Bcl-2 oligomerizes in membranes forming pores, but the extent of oligomerization and the size of the resulting pores are much smaller than that of Bax, supporting the model that Bcl-2 is a defective Bax.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • BH3 Interacting Domain Death Agonist Protein / metabolism*
  • Cell Membrane / drug effects
  • Cell Membrane / metabolism*
  • Chromatography, Gel
  • Cross-Linking Reagents / pharmacology
  • Kinetics
  • Liposomes / metabolism
  • Models, Biological
  • Molecular Weight
  • Mutation / genetics
  • Porins / metabolism*
  • Protein Binding / drug effects
  • Protein Multimerization* / drug effects
  • Proto-Oncogene Proteins c-bcl-2 / metabolism*
  • bcl-2-Associated X Protein / metabolism

Substances

  • BH3 Interacting Domain Death Agonist Protein
  • Cross-Linking Reagents
  • Liposomes
  • Porins
  • Proto-Oncogene Proteins c-bcl-2
  • bcl-2-Associated X Protein