Abstract
During transduction of an apoptotic (death) signal into the cell, there is an alteration in the permeability of the membranes of the cell's mitochondria, which causes the translocation of the apoptogenic protein cytochrome c into the cytoplasm, which in turn activates death-driving proteolytic proteins known as caspases1,2. The Bcl-2 family of proteins, whose members may be anti-apoptotic or pro-apoptotic, regulates cell death by controlling this mitochondrial membrane permeability during apoptosis3,4,5, but how that is achieved is unclear. Here we create liposomes that carry the mitochondrial porin channel (also called the voltage-dependent anion channel, or VDAC) to show that the recombinant pro-apoptotic proteins Bax and Bak accelerate the opening of VDAC, whereas the anti-apoptotic protein Bcl-xL closes VDAC by binding to it directly. Bax and Bak allow cytochrome c to pass through VDAC out of liposomes, but passage is prevented by Bcl-xL. In agreement with this, VDAC1-deficient mitochondria from a mutant yeast did not exhibit a Bax/Bak-induced loss in membrane potential and cytochrome c release, both of which were inhibited by Bcl-xL. Our results indicate that the Bcl-2 family of proteins bind to the VDAC in order to regulate the mitochondrial membrane potential and the release of cytochrome c during apoptosis.
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Acknowledgements
We thank E. Margoliash, G. Schatz and H. H. Schmid for antibodies against pigeon cytochrome c, yeast cytochrome c, and rat ANT, respectively; M. Forte for VDAC1-deficient yeast (M22-2), its wild type (M3), and human vdac1 cDNA; J. M. Hardwick for Bcl-xL mutant cDNAs; M. Colombini for the polyanion; T. Chittenden for the expression plasmid for the GST–Bak fusion protein; and N.Tsujimoto for editorial assistance. This study was supported by grants for Scientific Research on Priority Areas, for Center of Excellence Research and for Scientific Research, from the Ministry of Education, Science, Sports and Culture of Japan.
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Shimizu, S., Narita, M., Tsujimoto, Y. et al. Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature 399, 483–487 (1999). https://doi.org/10.1038/20959
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DOI: https://doi.org/10.1038/20959
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