Inhibition of pyruvate:ferredoxin oxidoreductase from Trichomonas vaginalis by pyruvate and its analogues. Comparison with the pyruvate decarboxylase component of the pyruvate dehydrogenase complex.
Pyruvate:ferredoxin oxidoreductase and the pyruvate dehydrogenase multi-enzyme complex both catalyse the CoA-dependent oxidative decarboxylation of pyruvate but differ in size, subunit composition and mechanism. Comparison of the pyruvate:ferredoxin oxidoreductase from the protozoon Trichomonas vaginalis and the pyruvate dehydrogenase component of the Escherichia coli pyruvate dehydrogenase complex shows that both are inactivated by incubation with pyruvate under aerobic conditions in the absence of co-substrates. However, only the former is irreversibly inhibited by incubation with hydroxypyruvate, and only the latter by incubation with bromopyruvate. Pyruvate:ferredoxin oxidoreductase activity is potently, but reversibly, inhibited by addition of bromopyruvate in the presence of CoA, and it is suggested that the mechanism involves formation of an adduct between CoA and bromopyruvate in the active site of the enzyme. It is proposed that both enzymes are inactivated by pyruvate through a mechanism involving oxidation of an enzyme-bound thiamin pyrophosphate/substrate adduct to form a tightly bound inhibitory species, possibly thiamin thiazolone pyrophosphate as hypothesized by Sumegi & Alkonyi.
Department of Biochemistry, University of Cambridge, U.K.
K P
KP
Williams
P F
PF
Leadlay
P N
PN
Lowe
D000818:Animals; D001665:Binding Sites; D002262:Carboxy-Lyases / Q000037:antagonists & inhibitors*; D003065:Coenzyme A / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology; D007658:Ketone Oxidoreductases / Q000037:antagonists & inhibitors*; D007700:Kinetics; D011767:Pyruvate Decarboxylase / Q000037:antagonists & inhibitors*; D011768:Pyruvate Dehydrogenase Complex / Q000037:antagonists & inhibitors*; D050743:Pyruvate Synthase; D011773:Pyruvates / Q000494:pharmacology*; D019289:Pyruvic Acid; D013835:Thiamine Pyrophosphate / Q000494:pharmacology; D014246:Trichomonas vaginalis / Q000201:enzymology*
D011768:Pyruvate Dehydrogenase Complex; D011773:Pyruvates; C017092:bromopyruvate; D019289:Pyruvic Acid; C012375:hydroxypyruvic acid; D007658:Ketone Oxidoreductases; D050743:Pyruvate Synthase; D002262:Carboxy-Lyases; D011767:Pyruvate Decarboxylase; D013835:Thiamine Pyrophosphate; D003065:Coenzyme A; D004229:Dithiothreitol