Pea stem mitochondria, resuspended in a KCl medium (de-energized mitochondria), underwent a swelling, as a consequence of K+ entry, that was inhibited by ATP. This inhibition was partially restored by GTP and diazoxide (K+ ATPchannel openers). In addition, glyburide and 5-hydroxydecanate (K+ ATP channel blockers) induced an inhibition of the GTP-stimulated swelling. Mitochondrial
swelling was inhibited by H2O2, but stimulated by NO. The same type of responses was also obtained in succinate-energized mitochondria. When the succinate-dependent transmembrane electrical potential (19) had reached a steady state, the addition of KCl induced a dissipation that was inhibited by H2O2and stimulated by NO. The latter stimulation was prevented by carboxy-PTIO, a NO scavenger. Phenylarsine oxide (a thiol oxidant) and NEM (a thiol blocker) stimulated the KCl-induced dissipation of19, while DTE prevented this effect in both cases. In addition, DTE transiently inhibited the NO-induced dissipation of19, but then it caused a more rapid collapse. These results,
therefore, show that the plant mitochondrial K+ ATP channel resembles that present in mammalian mitochondria and that it appears to be modulated by dithiol–disulfide interconversion, NO and H2O2.
The aperture of this channel was linked to the partial rupture of the outer membrane. The latter effect led to a release of cytochromec, thus suggesting that this release may be involved in the manifestation of programmed cell death.
