Studies on indoleamine 2,3-dioxygenase. I. Superoxide anion as substrate.
Indoleamine 2,3-dioxygenase purified to apparent homogeneity from rabbit intestine was inhibited by scavengers for superoxide anion such as superoxide dismutase and 1,2-dihydroxybenzene-3,5-disulfonic acid (Tiron). On the other hand, beta-carotene and 1,4-diazobicyclo-(2,2,2)-octane, scavengers for singlet oxygen, did not affect the enzyme activity significantly. The degree of inhibition of the dioxygenase by superoxide dismutase preparations from bovine erythrocytes, green peas, spinach leaves, and Escherichia coli paralleled that observed with these dismutase preparations on the aerobic reduction of cytochrome c by xanthine oxidase and its substrate. The pH profiles of the inhibition by dismutase of the dioxygenase and cytochrome c reduction were also similar and the maximal inhibition was observed around pH 10 in both cases. The degree of inhibition was not affected by the concentration of substrate but was a function of the concentration of dismutase. It was inversely related to the concentrations of the dioxygenase and its cofactors, ascorbic acid and methylene blue, both of which were required for maximum activity. Ascorbic acid could be replaced either by xanthine oxidase and its substrate, or by tetrabutylammonium superoxide prepared by electrolytic reduction of molecular oxygen, or by potassium superoxide. When limited amounts of superoxide anion were added to the reaction mixture containing a substrate amount of the dioxygenase, the ratio of the amount of superoxide anion added to that of the product formed was approximately unity both under aerobic and anaerobic conditions. Taken together, these findings indicate that superoxide anion, rather than molecular oxygen, is utilized as substrate by indoleamine 2,3-dioxygenase.
D014013:1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt / Q000494:pharmacology; D000588:Amines; D000818:Animals; D001391:Azo Compounds / Q000494:pharmacology; D002338:Carotenoids / Q000494:pharmacology; D002417:Cattle; D003300:Copper / Q000494:pharmacology; D004912:Erythrocytes / Q000201:enzymology; D004926:Escherichia coli / Q000201:enzymology; D006151:Guanosine / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007211:Indoles; D007422:Intestines / Q000201:enzymology*; D007700:Kinetics; D010105:Oxygenases / Q000378:metabolism*; D010545:Peroxides; D010944:Plants / Q000201:enzymology; D011817:Rabbits; D013045:Species Specificity; D013482:Superoxide Dismutase / Q000494:pharmacology; D013997:Time Factors
D000588:Amines; D001391:Azo Compounds; D007211:Indoles; D010545:Peroxides; D006151:Guanosine; D002338:Carotenoids; D014013:1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt; D003300:Copper; D010105:Oxygenases; D013482:Superoxide Dismutase