Enzymology of ubiquinone-utilizing electron transfer complexes in nonionic detergent.
The enzymology of isolated succinate: ubiquinone reductase and ubiquinone: cytochrome c reductase in nonionic detergents (alkyl polyoxyethylene derivatives) was studied. In the membrane the two multiprotein complexes and their hydrophobic substrates ubiquinone and dihydroubiquinone, are embedded in a common lipid bilayer. In detergent solutions the complexes are each inserted into micelles. Detergent micelles also serve as a solvent for the complexes hydrophobic substrates. As a consequence the isolated complexes are in a discontinuous phase with respect to their hydrophobic substrates and with respect to each other. Three types of assays were used. Firstly, single enzyme assays in which the hydrophobic substrates had to transfer from free micelles to the complex-bound micelles in order for enzyme reactions to occur. Secondly, assays in which the enzymic reactions were coupled to auxiliary nonenzymic reactions which rapidly converted the hydrophobic products back into substrates within the complex-bound micelle. Dichloroindophenol was used for the oxidation of dihydroubiquinone and dihydroduroquinone for the reduction of ubiquinone. Thirdly, assays in which the succinate: ubiquinone reductase reaction was coupled with the ubiquinone: cytochrome c reductase reaction. With the first type of assay, the kinetics of the substrate transfer reaction was dependent upon the type of detergent. In detergents with small polyoxyethylene head groups the transfer reactions were rate-limiting, and in detergents with large polyoxyethylene head groups the transfer reactions were fast and the enzymic reactions were rate-limiting...
D000818:Animals; D003579:Cytochrome Reductases / Q000378:metabolism*; D003574:Cytochrome c Group; D003902:Detergents / Q000494:pharmacology*; D004579:Electron Transport; D003576:Electron Transport Complex IV / Q000378:metabolism*; D007700:Kinetics; D008051:Lipid Bilayers; D008433:Mathematics; D008823:Micelles; D009097:Multienzyme Complexes / Q000378:metabolism*; D009206:Myocardium / Q000201:enzymology; D009245:NADH Dehydrogenase / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D010084:Oxidation-Reduction; D011808:Quinone Reductases / Q000378:metabolism*; D014451:Ubiquinone / Q000378:metabolism*
D003574:Cytochrome c Group; D003902:Detergents; D008051:Lipid Bilayers; D008823:Micelles; D009097:Multienzyme Complexes; D014451:Ubiquinone; D009247:NADH, NADPH Oxidoreductases; D003579:Cytochrome Reductases; D011808:Quinone Reductases; D009245:NADH Dehydrogenase; D003576:Electron Transport Complex IV