Location of protein(s) involved in oligomycin-induced inhibition of mitochondrial adenosinetriphosphatase near the outer surface of the inner membrane.
Mitoplasts, that is, mitochondria freed from their outer membranes, were prepared from pig heart. Sonication induced an inversion of these mitoplasts, giving inside-out vesicles. Added cytochrome c can be bound much better to mitoplasts than to sonicated vesicles; addition of trypsin increased adenosinetriphosphatase (ATPase) (ATP phosphohydrolase; EC 3.6.1.3) activity of sonicated vesicles without significantly affecting that of the mitoplasts. Since the site of fixation of cytochrome c was located on the outer side of the inner mitochondrial membrane and since the protein inhibitor of the mitochondrial ATPase is present on the inner face of the inner membrane and is very sensitive to trypsin, it can be concluded that mitoplasts are mainly oriented as normal mitochondria while sonicated vesicles are mainly inverted. Trypsin treatment can abolish the oligomycin sensitivity of ATPase activity of either mitoplasts or sonicated vesicles. However, trypsin induced the solubilization of the soluble F(1)-ATPase of sonicated vesicles while the ATPase activity remained with the mitoplasts after trypsin action. Therefore, trypsin destroyed the oligomycin effect by rupturing the liaison between F(1) and the membrane in sonicated vesicles. On the other hand, the effect of trypsin on mitoplasts must be attributed to the hydrolysis of a protein located near the outer surface of the inner membrane that is at least structurally involved in the oligomycin sensitivity of the ATPase complex.
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors*; D000818:Animals; D003574:Cytochrome c Group / Q000378:metabolism; D003593:Cytoplasm / Q000201:enzymology; D066298:In Vitro Techniques; D008566:Membranes / Q000201:enzymology; D008929:Mitochondria, Heart / Q000201:enzymology* / Q000648:ultrastructure; D009124:Muscle Proteins / Q000302:isolation & purification / Q000502:physiology*; D009840:Oligomycins / Q000494:pharmacology*; D013552:Swine; D014357:Trypsin / Q000494:pharmacology
D003574:Cytochrome c Group; D009124:Muscle Proteins; D009840:Oligomycins; D014357:Trypsin; D000251:Adenosine Triphosphatases