Glutathione adduct formation with microsomally activated metabolites of the pulmonary alkylating and cytotoxic agent, 3-methylindole.
Incubations with goat lung and liver microsomes were conducted to trap with exogenous glutathione (GSH) the electrophilic intermediate produced via cytochrome P-450-dependent metabolic activation of 3-methylindole (3MI). Microsomal incubation mixtures with [14C]3MI, a NADPH-generating system, and [3H]GSH produced a dual-labeled adduct which was isolated by reverse-phase high-performance liquid chromatography. Reactive 3MI intermediates were also trapped with cysteine. Adduct formation increased in proportion to the concentration of either thiol. Covalent binding of activated 3MI metabolites to microsomal protein was inversely related to adduct production. There were both qualitative and quantitative differences in the formation of GSH adducts by lung and liver microsomes. In the presence of 2 mM GSH, the adduct was produced at a rate of 1.8 nmol/mg protein/min by lung microsomes but only at 0.1 nmol/mg protein/min by hepatic microsomes. The addition of cytosolic fractions containing glutathione S-transferase activity increased GSH adduct formation by approximately 30%. These results support the view that electrophilic 3MI intermediates are trapped by conjugation with GSH, and that organ-selective toxicity is primarily due to much faster rates of cytochrome P-450 oxidation of 3MI in the lung than in the liver.
D000477:Alkylating Agents / Q000378:metabolism*; D000818:Animals; D001711:Biotransformation; D003577:Cytochrome P-450 Enzyme System / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D005260:Female; D005978:Glutathione / Q000378:metabolism*; D006041:Goats; D066298:In Vitro Techniques; D007211:Indoles / Q000378:metabolism*; D008168:Lung / Q000378:metabolism*; D008862:Microsomes, Liver / Q000378:metabolism*; D011485:Protein Binding; D012862:Skatole / Q000378:metabolism*
D000477:Alkylating Agents; D007211:Indoles; D003577:Cytochrome P-450 Enzyme System; D012862:Skatole; D005978:Glutathione