Mitochondrial release of pro-apoptotic proteins - Electrostatic interactions can hold cytochrome c but not Smac/DIABLO to mitochondrial membranes*
- Author(s)
- Uren, RT; Dewson, G; Bonzon, C; Lithgow, T; Newmeyer, DD; Kluck, RM;
- Details
- Publication Year 2005-01-21,Volume 280,Issue #3,Page 2266-2274
- Journal Title
- JOURNAL OF BIOLOGICAL CHEMISTRY
- Publication Type
- Journal Article
- Abstract
- A key step in the initiation of apoptosis is the release from the mitochondrial intermembrane space of cytochrome c and other pro-apoptotic proteins such as Smac/DIABLO, Omi/HtrA2, apoptosis-inducing factor (AIF), and endonuclease G (EndoG). Discrepancies have arisen, however, as to whether all these proteins are released in different systems. Our results suggest that failure to observe cytochrome c release may be due to the use of different buffers because after permeabilization by caspase-8 cleaved human Bid (tBid), cytochrome c dissociation from mitochondria was highly dependent on ionic strength and required 50 - 80 mM KCl, NaCl, or LiCl. In addition, mitochondria isolated from apoptotic cells using low ionic strength buffer bound a greater proportion of endogenous cytochrome c. In contrast to cytochrome c, Smac/DIABLO and Omi/HtrA2 were released independent of ionic strength, and AIF and EndoG behaved as if they are exposed to the intermembrane space but tethered to or within the inner membrane. AIF and EndoG were also not released by active caspases, which suggests their involvement in apoptosis may be limited. In summary, whereas tBid permeabilizes the outer membrane to cytochrome c, Smac/DIABLO, and Omi/HtrA2, the release of cytochrome c during apoptosis will be underestimated unless sufficient ionic strength is maintained to overcome the electrostatic association of cytochrome c with membranes.
- Publisher
- AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
- Keywords
- CELL-FREE APOPTOSIS; CASPASE ACTIVATION; OUTER-MEMBRANE; IONIC-STRENGTH; YEAST MITOCHONDRIA; EGG EXTRACTS; BINDING; BAX; CARDIOLIPIN; IMPORT
- Publisher's Version
- https://doi.org/10.1074/jbc.M411106200
- Terms of Use/Rights Notice
- Refer to copyright notice on published article.
Creation Date: 2005-01-21 12:00:00