NN'-dicyclohexylcarbodi-imide-sensitivity of bovine heart mitochondrial NADH: ubiquinone oxidoreductase. Inhibition of activity and binding to subunits.
Dicyclohexylcarbodi-imide (DCCD) inhibition of NADH: ubiquinone oxidoreductase was studied in submitochondrial particles and in the isolated form, together with the binding of the reagent to the enzyme. DCCD inhibited the isolated enzyme in a time- and concentration-dependent manner. Over the concentration range studied, a maximum inhibition of 85% was attained within 60 min. The time course for the binding of DCCD to the enzyme was similar to that of activity inhibition. The NADH:ubiquinone oxidoreductase activity of the submitochondrial particles was also sensitive to DCCD, and the locus of binding of the inhibitor was studied by subsequent resolution of the enzyme into subunit polypeptides. Only two subunits (molecular masses 13.7 and 21.5 kDa) were labelled by [14C]DCCD, whereas, when the enzyme in its isolated form was treated with [14C]DCCD, six subunits (13.7, 16.1, 21.5, 39, 43 and 53 kDa) were labelled. Comparison with the subunit labelling of F1F0-ATPase and ubiquinol:cytochrome c oxidoreductase indicated that the labelling pattern of NADH:ubiquinone oxidoreductase, and enzyme complex with a multitude of subunits, is unique and not due to contamination by other inner-membrane proteins. The correlation between the electron- and proton-transport functions and the DCCD-binding components remains to be established.
D000818:Animals; D002234:Carbodiimides / Q000494:pharmacology*; D002250:Carbon Radioisotopes; D002417:Cattle; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology*; D046911:Macromolecular Substances; D008929:Mitochondria, Heart / Q000187:drug effects / Q000201:enzymology*; D016660:NAD(P)H Dehydrogenase (Quinone); D011485:Protein Binding; D011808:Quinone Reductases / Q000037:antagonists & inhibitors*; D013367:Submitochondrial Particles / Q000187:drug effects / Q000201:enzymology
D002234:Carbodiimides; D002250:Carbon Radioisotopes; D046911:Macromolecular Substances; D004024:Dicyclohexylcarbodiimide; D016660:NAD(P)H Dehydrogenase (Quinone); D011808:Quinone Reductases