The active form of cytochrome c oxidase: effects of detergent, the intact membrane, and radiation inactivation.
Cytochrome oxidase is a multisubunit, intrinsic membrane protein with a complex function that includes oxidation of cytochrome c, reduction of oxygen and generation of a membrane potential. To clarify the relationship of its normal function to protein and membrane structure, we have examined the kinetic behavior of rat liver cytochrome oxidase in the intact inner mitochondrial membrane and in detergent solubilized states. Dissolution of rat liver mitochondrial membranes alters the kinetic parameters of the oxidase in a manner dependent in part on the dispersing agent, and characterized by a large increase in maximal activity which is not attributable to exposure of more oxidase or diminished affinity for cytochrome c. The most profound effect of solubilization of the membrane is seen on the low affinity reaction of cytochrome c, suggesting that the electron transfer pathway from this site to oxygen is sensitive to alterations in hydrophobic interactions within the oxidase. Purified rat liver and beef heart oxidase exists predominantly in a monodisperse, 300 kilodalton form in laurylmaltoside (Rosevear et al., 1980). However, a smaller, 130 kd species that exhibits high turnover rates equal to the 300 kd form is detected in some beef heart preparations, implying that the dimer may not be essential for high activity. Radiation inactivation studies on purified oxidase reveal a molecular weight for the functional unit of approximately 70 kd. It is concluded that less than a complete set of subunits may be sufficient for both normal binding of cytochrome c and rapid electron transfer to oxygen.
D000818:Animals; D002417:Cattle; D003902:Detergents / Q000494:pharmacology; D004579:Electron Transport; D003576:Electron Transport Complex IV / Q000502:physiology* / Q000528:radiation effects; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000201:enzymology*; D007700:Kinetics; D008567:Membranes, Artificial; D008930:Mitochondria, Liver / Q000201:enzymology; D008954:Models, Biological; D051381:Rats
D003902:Detergents; D008567:Membranes, Artificial; D003576:Electron Transport Complex IV