In rat liver mitochondria suspended in KC1 medium, oligomycin interfered with the effect of uncouplers on energy conservation. It antagonized the effect of uncouplers that are weak acids (2,4-dinitrophenol etc.), but enhanced that of the lipid-penetrating cation NN-dimethyl-N'N'-dibenzylammonium. Oligomycin caused none of the above effects when Br or NO/3 was substituted for C1 as the major anionic species in the assay medium. The concentration of oligomycin that exerted the above-mentioned effects was lower than that necessary for the inhibition of energy transfer, but was in the range that induced C1 permeation through the cristae membrane. The possible connexion between the effect of oligomycin on C1 permeation and its interference with the action of uncouplers is discussed.

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