Bax-induced Cytochrome C Release from Mitochondria Is Independent of the Permeability Transition Pore but Highly Dependent on Mg2+ Ions

doi:10.1083/jcb.143.1.217

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© The Rockefeller University Press, 0021-9525/1998//217 $5.00
The Journal of Cell Biology, Volume 143, Number 1, , 1998 217-224

Regular Articles

Bax-induced Cytochrome C Release from Mitochondria Is Independent of the Permeability Transition Pore but Highly Dependent on Mg²⁺ Ions

Robert Eskes^*, Bruno Antonsson^*, Astrid Osen-Sand^*, Sylvie Montessuit^*, Christoph Richter, Rémy Sadoul^*, Gonzalo Mazzei^*, Anthony Nichols^*, and Jean-Claude Martinou^*

^* Serono Pharmaceutical Research Institute, 1228 Plan-les-Ouates, Geneva, Switzerland; and ETH Laboratory of Biochemistry, 8092 Zurich, Switzerland

Bcl-2 family members either promote or repress programmed celldeath. Bax, a death-promoting member, is a pore-forming, mitochondria-associated protein whose mechanism of action is still unknown. Duringapoptosis, cytochrome C is released from the mitochondria intothe cytosol where it binds to APAF-1, a mammalian homologueof Ced-4, and participates in the activation of caspases. Therelease of cytochrome C has been postulated to be a consequenceof the opening of the mitochondrial permeability transitionpore (PTP). We now report that Bax is sufficient to trigger the release of cytochrome C from isolated mitochondria. Thispathway is distinct from the previously described calcium-inducible,cyclosporin A–sensitive PTP. Rather, the cytochrome Crelease induced by Bax is facilitated by Mg²⁺ and cannot beblocked by PTP inhibitors. These results strongly suggest theexistence of two distinct mechanisms leading to cytochromeC release: one stimulated by calcium and inhibited by cyclosporinA, the other Bax dependent, Mg²⁺ sensitive but cyclosporininsensitive.

Key Words: apoptosis • mitochondria • Bax • cytochrome C • permeability transition

Abbreviations used in this paper: BKA, bongkrekic acid; CsA, cyclosporin A; MB, mitochondrial buffer; PTP, permeability transition pore; PT, permeability.

Address all correspondence to J.-C. Martinou, Serono Pharmaceutical Research Institute, 14 Chemin des Aux, 1228 Plan les Ouates,Geneva, Switzerland. Tel.: (41) 22 706 9822. Fax: (41) 22 7946965. E-mail: jean-claude.martinou.ch_gva03{at}serono.com

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