Inhibition of mitochondrial NADH:ubiquinone oxidoreductase by ethoxyformic anhydride.
The NADH:ubiquinone, but not the NADH:ferricyanide, reductase activity of mitochondrial complex I (NADH:ubiquinone oxidoreductase) is inhibited by incubation of the enzyme at pH 6.0 and 0 degree C with ethoxyformic anhydride (EFA), and the inhibition is partially reversed by subsequent incubation of EFA-treated complex I with hydroxylamine. These results and spectral changes of EFA-treated complex I in the u.v. region are consistent with modification of essential histidyl or tyrosyl residues between the primary NADH dehydrogenase and the site of ubiquinone reduction. Treatment of complex I with EFA in the presence of high concentrations of Seconal or Demerol did not protect against EFA inactivation, suggesting that the site of EFA modification may not be the same as the inhibiton sites of Seconal and Demerol. However, the presence of NADH during incubation of complex I with EFA greatly enhanced the inhibition rate, indicating that the reduced conformation of complex I is more susceptible to attack by EFA.
D000818:Animals; D002417:Cattle; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology*; D004579:Electron Transport; D005561:Formates / Q000494:pharmacology*; D006639:Histidine / Q000037:antagonists & inhibitors; D019811:Hydroxylamine; D006898:Hydroxylamines / Q000494:pharmacology; D007700:Kinetics; D008614:Meperidine / Q000494:pharmacology; D008929:Mitochondria, Heart / Q000201:enzymology*; D009243:NAD / Q000494:pharmacology; D016660:NAD(P)H Dehydrogenase (Quinone); D009247:NADH, NADPH Oxidoreductases / Q000037:antagonists & inhibitors*; D011808:Quinone Reductases / Q000037:antagonists & inhibitors*; D014443:Tyrosine / Q000037:antagonists & inhibitors
D005561:Formates; D006898:Hydroxylamines; D009243:NAD; D019811:Hydroxylamine; D014443:Tyrosine; D006639:Histidine; D008614:Meperidine; D009247:NADH, NADPH Oxidoreductases; D016660:NAD(P)H Dehydrogenase (Quinone); D011808:Quinone Reductases; D004047:Diethyl Pyrocarbonate