Inactivation of succinate-ubiquinone reductase in substrate mixture.
Succinate-ubiquinone reductase plays an important role in the respiratory chain. Previous work showed that preparation of succinate-ubiquinone reductase was relatively stable. Though the enzyme catalysis has been extensively studied, the inactivation of succinate-ubiquinone reductase has never been reported. In the present study, the kinetic theory of the substrate reaction of irreversible inhibition described by Tsou (Adv. Enzymol. Relat. Areas Mol. Biol. 61 (1988) 381-436) was applied to study the course of an unexpected slow inactivation of succinate-ubiquinone reductase in the substrate assay mixture containing different concentrations of substrates, succinate and 2,6-dichloroindophenol. The results showed that the inactivation of succinate-ubiquinone reductase in the substrate mixture is a first order reaction. The inactivation rate decreased with increasing concentration of succinate. The values of the micro rate constants for free and succinate bound enzyme were 0.22 +/- 0.01 and 0.052 +/- 0.002 min-1, respectively. Binding with 2-thenoyl-trifluroacetone, a inhibitor specially for the quinone binding site, slowed down the inactivation. However, the rate of inactivation did not change with increasing 2,6-dichloroindophenol concentration. The study showed that succinate-ubiquinone reductase was irreversibly inactivated in the substrate mixture. The results suggest that the inactivation was not due to dilution or dissociation of the enzyme, nor to complete usage of the substrate, inhibition of the yielded product or some possible trace component in the substrate mixture, nor to modification of the essential thiol group in the succinate binding site of succinate-ubiquinone reductase. The enzyme became more stable after binding with succinate.
Yang
Y
Y
School of Life Science and Engineering, Tsinghua University, Beijing, China.
Xu
J X
JX
Zhou
H M
HM
D015086:2,6-Dichloroindophenol / Q000737:chemistry / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D042963:Electron Transport Complex II; D007700:Kinetics; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors* / Q000737:chemistry; D009206:Myocardium / Q000201:enzymology*; D010088:Oxidoreductases / Q000037:antagonists & inhibitors* / Q000737:chemistry; D013379:Substrate Specificity; D013385:Succinate Dehydrogenase / Q000037:antagonists & inhibitors* / Q000737:chemistry; D019802:Succinic Acid / Q000737:chemistry / Q000494:pharmacology*
D009097:Multienzyme Complexes; D019802:Succinic Acid; D015086:2,6-Dichloroindophenol; D010088:Oxidoreductases; D042963:Electron Transport Complex II; D013385:Succinate Dehydrogenase