Inhibition of mouse glutathione transferases and glutathione peroxidase II by dicumarol and other ligands.
Dicumarol, often used as a specific inhibitor of DT diaphorase (NAD(P)H:(quinone-acceptor) oxidoreductase; EC 1.6.99.2), was found to potently inhibit GSH transferases (EC 2.5.1.18). Dicumarol exhibited an IC50 of 11 microM in inhibiting the conjugation of 1-chloro-2,4-dinitrobenzene (50 microM) by GSH transferase GT-8.7, the major hepatic class mu isoenzyme of CD-1 mice. The activities of GT-8.7 and of the class pi isoenzyme, GT-9.0, toward a carcinogenic substrate, 4-nitroquinoline 1-oxide (100 microM), were inhibited by dicumarol with IC50 values of 14 and 9 microM, respectively. Dicumarol also affected GSH peroxidase II activity, inhibiting the reduction of cumene hydroperoxide by GT-10.6, the predominant class alpha GSH transferase of mouse liver, with an IC50 of 14 microM. GSH peroxidase I (EC 1.11.1.9) and GSH peroxidase II activities were resolved by chromatography of liver and testis cytosols. While inhibiting GSH peroxidase II with IC50 of 9-10 microM, dicumarol did not affect the activity of the selenoenzyme, GSH peroxidase I. Whereas several other non-substrate ligands were more potent inhibitors of 1-chloro-2,4-dinitrobenzene conjugation, dicumarol effectively inhibited GSH transferase and GSH peroxidase II activities in the range of dicumarol concentrations frequently used for detection of DT diaphorase action. These results indicate that physiological consequences resulting from the use of supramicromolar concentrations of dicumarol should not be interpreted in terms of DT diaphorase inhibition alone.
Mays
J B
JB
Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock 72205-7199.
Benson
A M
AM
D015112:4-Nitroquinoline-1-oxide / Q000378:metabolism; D000818:Animals; D001555:Benzene Derivatives / Q000378:metabolism; D003600:Cytosol / Q000187:drug effects / Q000201:enzymology; D001728:Dicumarol / Q000494:pharmacology*; D004137:Dinitrochlorobenzene / Q000378:metabolism; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors*; D007527:Isoenzymes / Q000037:antagonists & inhibitors*; D008099:Liver / Q000187:drug effects / Q000201:enzymology; D008297:Male; D051379:Mice; D010084:Oxidation-Reduction / Q000187:drug effects; D010544:Peroxidases / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D013737:Testis / Q000187:drug effects / Q000201:enzymology
D001555:Benzene Derivatives; D004137:Dinitrochlorobenzene; D007527:Isoenzymes; D015112:4-Nitroquinoline-1-oxide; D001728:Dicumarol; D010544:Peroxidases; C027585:glutathione peroxidase II; D005982:Glutathione Transferase; C007164:cumene hydroperoxide