In this study we applied the potential-sensitive fluorescent dye indocyanine to measure the initial rate of membrane potential development in liposomes containing reconstituted cytochrome c oxidase from bovine heart. A linear relationship was found between the initial rate of membrane potential development, related to the H+/e(-)-stoichiometry, and the rate of electron flow at various substrate concentrations. By increasing the salt concentration a decrease of the initial rate of membrane potential development was found. A similar dependence on the salt concentration was also found for the membrane potential under steady state conditions. The data further demonstrate variation of the H+/e(-)-stoichiometry in cytochrome c oxidase depending on the salt concentration in the buffer.
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