Generation of Transmembrane Electrical Potential during NADH Oxidation via the External Pathway and the Fatty Acid Uncoupling Effect after Transient Opening of the Ca²⁺-Dependent Cyclosporin A-Sensitive Pore in Liver Mitochondria

M. E. Bodrova, V. I. Dedukhova, and E. N. Mokhova^*

Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, 119899 Russia; fax: (095) 939-3181; E-mail: mokhova@genebee.msu.su

^* To whom correspondence should be addressed.

Received June 8, 1999; Revision received October 21, 1999

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The effects of transient pore opening on generation of the transmembrane gradient of electrical potential across the inner mitochondrial membrane (DeltaPsi) induced by NADH oxidation through the external pathway as well as on the uncoupling effect of fatty acids were studied. The pore opening was monitored by changes in the DeltaPsi value. The cycle of pore opening/closing was found to have only an insignificant effect on the sensitivity of DeltaPsi to fatty acid uncoupling. Once this cycle is over, NADH oxidation in the presence of exogenous cytochrome c results in generation of DeltaPsi. In the absence of cytochrome c, the generation of DeltaPsi induced by oxidation of exogenous NADH is observed if the incubation medium pH has been decreased from 7.4 to 7.0. The generation of DeltaPsi was inhibited by cyclosporin A. In isotonic salt medium containing 125 mM KCl, the maximum level of DeltaPsi generated by exogenous NADH after the cycle of pore opening/closing was significantly lower than the maximum level of DeltaPsi generated in hypotonic incubation medium. The data obtained in this work suggest that the cycle of pore opening/closing has little if any effect on the energy coupling in liver mitochondria, whereas the external pathway of NADH oxidation activated by this cycle may support the energy-dependent functions of liver mitochondria.

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KEY WORDS: mitochondrial permeability transition, transmembrane electrical potential difference, exogenous NADH, cytochrome c, fatty acids, liver mitochondria

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