- 作者: Maria L. R. Lim May-Ghee Lum, Tania M. Hansen, Xavier Roucou, Phillip Nagley
- 作者服務機構: Department of Biochemistry and Molecular Biology, Monash University, Melbourne, Vic., Australia
- 中文摘要: --
- 英文摘要: of cyt c-GFP fusions and the determination of the charac-teristics of the putative outer membrane pore throughwhich cyt c and other mitochondrial proteins with pro-apoptotic functions may pass. The richness of this fieldrelating to mitochondria and cell death is brought out byconsideration of studies carried out in mammal ian andyeast cells.Mitochondria play key roles in apoptosis, a central stepbeing the release of cytochrome c (cyt c) across the outermitochondrial membrane into the cytoplasm. We reviewthis process in terms of the influences that induce mito-chondria to release cyt c, the possible mechanisms ofsuch release and the downstream consequences for cas- pase activation. The contributions of members of the Bcl-2 family in regulating mitochondrial activities relevant toapoptotic signaling are considered. Antiapoptotic mem-bers, such as Bcl-2 itself, are antagonistic to other familymembers, which prominently include Bax amongst ahost of other proapoptotic proteins homologous to Bcl-2. Focus is placed on technical methods of determining cytc release, which encompass cell fractionation,biochem-istry, immunochemistry and confocal microscopy [in-cluding observations of release in real time using cyt c-green fluorescent protein (GFP) fusion proteins]. Theadvantages and potential pitfalls of the various ap-proaches are discussed, with some emphasis on the use
- 中文關鍵字: --
- 英文關鍵字: Mitochondria, Cytochromec, Apoptosis · Green fluorescent protein, Bcl-2, Bax, Caspases, Confocal microscopy, Membrane potential, Signaling, Yeast, Mammalian Cells