Bcl-2 Prevents Bax Oligomerization in the Mitochondrial Outer Membrane

Valery Mikhailov, Margarita Mikhailova, Donna J. Pulkrabek, Zheng Dong, Manjeri A. Venkatachalam, Pothana Saikumar

Research output: Contribution to journalArticle

266 Scopus citations

Abstract

ATP depletion results in Bax translocation from cytosol to mitochondria and release of cytochrome c from mitochondria into cytosol in cultured kidney cells. Overexpression of Bcl-2 prevents cytochrome c release, without ameliorating ATP depletion or Bax translocation, with little or no association between Bcl-2 and Bax as demonstrated by immunoprecipitation (Saikumar, P., Dong, Z., Patel, Y., Hall, K., Hopfer, U., Weinberg, J. M., and Venkatachalam, M. A. (1998) Oncogene 17, 3401-3415). Now we show that translocated Bax forms homooligomeric structures, stabilized as chemical adducts by bifunctional cross-linkers in ATP-depleted wild type cells, but remains monomeric in Bcl-2-overexpressing cells. The protective effects of Bcl-2 did not require Bcl-2/Bax association, at least to a degree of proximity or affinity that was stable to conditions of immunoprecipitation or adduct formation by eight cross-linkers of diverse spacer lengths and chemical reactivities. On the other hand, nonionic detergents readily induced homodimers and heterodimers of Bax and Bcl-2. Moreover, associations between translocated Bax and the voltage-dependent anion channel protein or the adenine nucleotide translocator protein could not be demonstrated by immunoprecipitation of Bax, or by using bifunctional cross-linkers. Our data suggest that the in vivo actions of Bax are at least in part dependent on the formation of homo-oligomers without requiring associations with other molecules and that Bcl-2 cytoprotection involves mechanisms that prevent Bax oligomerization.

Original languageEnglish (US)
Pages (from-to)18361-18374
Number of pages14
JournalJournal of Biological Chemistry
Volume276
Issue number21
DOIs
StatePublished - Jan 25 2001
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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    Mikhailov, V., Mikhailova, M., Pulkrabek, D. J., Dong, Z., Venkatachalam, M. A., & Saikumar, P. (2001). Bcl-2 Prevents Bax Oligomerization in the Mitochondrial Outer Membrane. Journal of Biological Chemistry, 276(21), 18361-18374. https://doi.org/10.1074/jbc.M100655200