This study was designed to determine which enzyme activities were first impaired in mitochondria exposed to 2,2′-azobis-(2-amidinopropane) dihydrochloride (AAPH), a known radical initiator. EPR spin-trapping revealed generation of reactive oxygen species although malondialdehyde formation remained very low. With increasing AAPH concentrations, State-3 respiration was progressively depressed with unaltered ADP/O ratios. A top-down approach demonstrated that alterations were located at the phosphorylation level. As shown by inhibitor titrations, ATP/ADP translocase activity was unaffected in the range of AAPH concentrations used. In contrast, AAPH appeared to exert a deleterious effect at the level of F1FO-ATPase, comparable with dicyclohexylcarbodi-imide, which alters FO proton channel. A comparison of ATP hydrolase activity in uncoupled and broken mitochondria reinforced this finding. In spite of its pro-oxidant properties, AAPH was shown to act as a dose-dependent inhibitor of cyclosporin-sensitive permeability transition initiated by Ca2+, probably as a consequence of its effect on F1FO-ATPase. Resveratrol, a potent antiperoxidant, completely failed to prevent the decrease in State-3 respiration caused by AAPH. The data suggest that AAPH, when used under mild conditions, acted as a radical initiator and was capable of damaging F1FO-ATPase, thereby slowing respiratory chain activity and reducing mitochondrial antioxidant defences.
F1F0-ATPase, early target of the radical initiator 2,2′-azobis-(2-amidinopropane) dihydrochloride in rat liver mitochondria in vitro
- Views Icon Views
- PDF LinkPDF
- Share Icon Share
- Cite Icon Cite
Frédéric BEAUSEIGNEUR, Marc GOUBERN, Marie-France CHAPEY, Joseph GRESTI, Catherine VERGELY, Marcelline TSOKO, Jean DEMARQUOY, Luc ROCHETTE, Pierre CLOUET; F1F0-ATPase, early target of the radical initiator 2,2′-azobis-(2-amidinopropane) dihydrochloride in rat liver mitochondria in vitro. Biochem J 1 December 1996; 320 (2): 571–576. doi: https://doi.org/10.1042/bj3200571
Download citation file: