Kinetics of modification of the mitochondrial succinate-ubiquinone reductase by 5,5'-dithiobis-(2-nitro-benzoic acid).
The kinetic theory of the substrate reaction during modification of enzyme activity previously described by Tsou [Tsou (1988), Adv. Enzymol. Relat. Areas Mol. Biol. 61, 381-436] has been applied to a study of the kinetics of the course of inactivation of the mitochondrial succinate-ubiquinone reductase by 5,5'-dithiobis-(2-nitro-benzoic acid) (DTNB). The results show that the inactivation of this enzyme by DTNB is a conformation-change-type inhibition which involves a conformational change of the enzyme before inactivation. The microscopic rate constants were determined for the reaction of the inactivator with the enzyme. The presence of the substrate provides marked protection of this enzyme against inactivation by DTNB. The modification reaction of the enzyme using DTNB was shown to follow a triphasic course by following the absorption at 412 nm. Among these reactive thiol groups, the fast-reaction thiol group is essential for the enzyme activity. The results suggest that the essential thiol group is situated at the succinate-binding site of the mitochondrial succinate-ubiquinone reductase.
Yang
Y
Y
School of Life Science and Engineering, Tsinghua University, Beijing, China.
Wang
H R
HR
Xu
J X
JX
Zhou
H M
HM
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology*; D042963:Electron Transport Complex II; D007700:Kinetics; D008928:Mitochondria / Q000201:enzymology; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010088:Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013385:Succinate Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013386:Succinates / Q000378:metabolism; D019802:Succinic Acid; D013439:Sulfhydryl Reagents / Q000494:pharmacology*
D009097:Multienzyme Complexes; D013386:Succinates; D013439:Sulfhydryl Reagents; D004228:Dithionitrobenzoic Acid; D019802:Succinic Acid; D010088:Oxidoreductases; D042963:Electron Transport Complex II; D013385:Succinate Dehydrogenase