Induction of the permeability transition and cytochrome c release by 15-deoxy-Δ12,14-prostaglandin J2 in mitochondria

The electrophilic lipid 15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)) is known to allow adaptation to oxidative stress in cells at low concentrations and apoptosis at high levels. The mechanisms leading to adaptation involve the covalent modification of regulatory proteins, such as Keap1, and augmentation of antioxidant defences in the cell. The targets leading to apoptosis are less well defined, but mitochondria have been indirectly implicated in the mechanisms of cell death mediated by electrophilic lipids. To determine file potential of electrophilic cyclopentenones to induce pro-apoptotic effects in the mitochondrion, we used isolated liver mitochondria and demonstrated that 15d-PGJ(2) promotes Ca2+- induced mitochondrial swelling and cytochrome c release. The mechanisms involved tire consistent with direct modification of protein thiols in file mitochondrion, rather than secondary formation of reactive oxygen species or lipid peroxidation. Using protcomic analysis in combination with biotinylated 15d-PGJ(2), we were able to identify 17 potential targets of the electrophile-responsive proteome in isolated liver mitochondria. Taken together, these results Suggest that electrophilic lipid oxidation products can target a sub-proteome in mitochondria, and this in turn results in file transduction of the electrophilic stimulus to the cell through cytochroine c release.