Uncoupler-inhibitor titrations of ATP-driven reverse electron transfer in isolated rat-liver mitochondria

Abstract

Uncoupler-inhibitor titrations of ATP-driven reverse electron transfer across the first site of the respiratory chain were performed in isolated rat-liver mitochondria, and the experimental results were compared with the predictions of a simple delocalised chemiosmotic mechanism. The rates of ATP hydrolysis ( J p) and reverse electron transfer (− J 0) were measured at different uncoupler (S-13) concentrations, either in the absence or in the presence of rotenone. When the rates − J 0 and J p measured at different uncoupler concentrations were expressed as percentages of the activity at zero uncoupler concentration, it was found that the efficiency of S-13 to uncouple the reverse electron transfer and to stimulate ATP hydrolysis was not significantly changed upon partial inhibition with rotenone. These results are in contrast with data from a study of uncoupler-inhibitor titrations in submitochondrial particles published previously, in which a higher effectiveness of several uncouplers to inhibit ATP-driven reverse electron transfer was observed in the presence of rotenone.