Inhibition of ceruloplasmin and other copper oxidases by thiomolybdate.
The dietary antagonism between copper and molybdate salts prompted a study of the inhibition of copper enzymes by thiomolybdate (TM). TM strongly inhibited the oxidase activity of five copper oxidase with I50% values in the 1-5 microM range. The mechanism of the TM effect on the copper oxidase, ceruloplasmin (Cp) (E.C. 1.16.3.1), was studied in detail. In Vmax vs. E plots, TM gave parallel data suggesting irreversibility but a large number of TM molecules per Cp were required. The inhibition of Cp by TM could not be reversed by dialysis. Isolation of TM-inhibited Cp on Sephadex G-10 did not yield any active Cp molecules. Cu(II) did not restore any inhibited oxidase activity. Gel electrophoresis supported the covalent binding of Cp by TM without any extensive change in protein structure. EPR results confirmed that Cu(II) is reduced to Cu(I) after reaction with TM. However, the Mo(VI) in MoS4(2-) did not change in oxidation number. Analysis of the TM-Cp compound accounted for all six Cu atoms as found in native Cp. The data suggest the covalent binding of sulfide to Cp copper. TM also inhibited the activity of ascorbate oxidase, cytochrome oxidase, superoxide dismutase, and tyrosinase. However, no inhibition of carbonic anhydrase, a zinc enzyme, was observed at 1 mM TM.
D000818:Animals; D001204:Ascorbate Oxidase / Q000037:antagonists & inhibitors; D002417:Cattle; D002570:Ceruloplasmin / Q000037:antagonists & inhibitors*; D003576:Electron Transport Complex IV / Q000037:antagonists & inhibitors; D006801:Humans; D066298:In Vitro Techniques; D008982:Molybdenum / Q000494:pharmacology*; D014442:Monophenol Monooxygenase / Q000037:antagonists & inhibitors; D010088:Oxidoreductases / Q000037:antagonists & inhibitors*; D013482:Superoxide Dismutase / Q000037:antagonists & inhibitors; D013552:Swine
D008982:Molybdenum; C020809:tetrathiomolybdate; D010088:Oxidoreductases; D001204:Ascorbate Oxidase; D014442:Monophenol Monooxygenase; D013482:Superoxide Dismutase; C024748:copper oxidase; D002570:Ceruloplasmin; D003576:Electron Transport Complex IV