Activation of beef-heart cytochrome c oxidase by cardiolipin and analogues of cardiolipin.
Beef-heart cytochrome c oxidase lacking endogenous lipids can be prepared by cholate-mediated exchange with dimyristoylphosphatidylcholine (Powell, G. L., Knowles, P. F. and Marsh, D. (1985) Biochim. Biophys. Acta 816, 191-194). These preparations retained practically no endogenous cardiolipin (less than 0.19 mol cardiolipin per mol of oxidase) but in Tween 80 they retained unaltered electron transport activity. Resupplementation of the dimyristoylphosphatidylcholine-substituted cytochrome oxidase with cardiolipin and cardiolipin analogues with different numbers of acyl chains or with a methylated headgroup enhanced the activity of the reconstituted enzyme to an extent dependent on the structure of the cardiolipin derivative. The Eadie-Hofstee plot showed biphasic kinetic behavior for all reconstituted preparations, even those completely lacking cardiolipin. This biphasic substrate dependence of the kinetics was simulated using the model of Brzezinski, P. and Malmström, B. G. (Proc. Natl. Acad. Sci. USA 83 (1986) 4282-4286), which implicates two interconverting enzyme conformations in the proton transport step. The activation of cytochrome c oxidase by the cardiolipin analogues could be explained in terms of an electrostatic enhancement of the surface concentrations of both cytochrome c and protons, and a facilitated interconversion between the two enzyme conformations.
Abramovitch
D A
DA
Department of Biological Sciences, Clemson University, SC 29634-1903.
Marsh
D
D
Powell
G L
GL
D000818:Animals; D002308:Cardiolipins / Q000494:pharmacology*; D002417:Cattle; D004134:Dimyristoylphosphatidylcholine / Q000494:pharmacology; D003576:Electron Transport Complex IV / Q000378:metabolism*; D004789:Enzyme Activation; D007700:Kinetics; D008929:Mitochondria, Heart / Q000201:enzymology*; D008962:Models, Theoretical; D010084:Oxidation-Reduction; D013329:Structure-Activity Relationship
D002308:Cardiolipins; D003576:Electron Transport Complex IV; D004134:Dimyristoylphosphatidylcholine