Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is a unique antioxidant enzyme that markedly reduces lipid hydroperoxide generated in biomembranes. Overexpression of mitochondrial PHGPx potentially suppresses the release of cytochrome c (cyt. c) from mitochondria and apoptosis. The hydroperoxide level in mitochondria was elevated in 2-deoxyglucose (2DG)-induced apoptosis, but not in apoptosis-resistant cells in which mitochondrial PHGPx was overexpressed. From studies of the overexpression of PHGPx, the generation of hydrogen peroxide and lipid hydroperoxide in mitochondria might be important triggers of apoptosis. In particular lipid hydroperoxide could be involved in the initiation of cyt. c liberation from mitochondria in 2DG-induced apoptosis since lipid hydroperoxide is a primary substrate for PHGPx. The release of cyt. c from mitochondria is an important proapoptotic signal in the mitochondrial death pathway. Several reports demonstrated the reactive oxgen species could be involved in cyt. c liberation, although its mechanism is still unknown. Cardiolipin (CL), which exclusively locates in the innermembrane of mitochondria, shows strong affinity for cyt. c is required for the adenine nucleotide translocator (ANT) that cotnrols the opening and closing of the permeability transition pore. Association of cyt. c with CL is lost upon peroxidation. CL hydroperoxide (CLOOH), in contrast to CL, does not bind to cyt. c. Furthermore, CLOOH can open the permeability transion pore by the inactivation of ANT. These previous results suggest that mitochondrial PHGPx inhibits the release of cyt. c from mitochondria by the scavenging CLOOH and could prevent apoptosis.