Effect of cytosolic components on the metabolism of the hydrazide iproniazid.
The effects of thiols, such as glutathione (GSH), and the cytosolic glutathione S-transferases on the microsomal metabolism of the hydrazide iproniazid to hydrocarbon products were investigated. Thiol compounds stimulated propane production and depressed propylene production. Addition of preparations of cytosolic proteins to the microsomal reaction mixtures in the presence of GSH depressed production of propane by more than 80% and propylene by 50% compared to the GSH-mediated reaction. The purified glutathione S-transferases A and B were most potent in eliciting this effect; isozymes AA, C, and E had little or no effect on hydrocarbon production. Further, a mixture of these purified isozymes in the concentrations known to exist in cytosol affected hydrocarbon production in a manner similar to cytosol. Experiments performed with isolated hepatocytes and an inhibitor of these cytosolic enzymes further supported the involvement of these enzymes in altered hydrocarbon production. These isozymes were subsequently shown to catalyze the formation of a GSH conjugate, S-(2-propyl)glutathione. The decreases in hydrocarbon production by microsomes in the presence of the glutathione S-transferases and GSH were accompanied by production of slightly larger amounts of conjugate. These data indicate that the cytosolic glutathione S-transferases interact with an oxidative microsomal metabolite of iproniazid to enzymatically produce an S-(2-propyl)glutathione conjugate and thus prevent formation of a reactive species which would otherwise chemically decompose to yield hydrocarbons or to covalently bind to cellular macromolecules.
D000818:Animals; D003600:Cytosol / Q000378:metabolism; D005978:Glutathione / Q000031:analogs & derivatives / Q000138:chemical synthesis; D005982:Glutathione Transferase / Q000302:isolation & purification / Q000378:metabolism; D007490:Iproniazid / Q000378:metabolism*; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry; D008862:Microsomes, Liver / Q000378:metabolism; D051381:Rats; D011916:Rats, Inbred F344; D011919:Rats, Inbred Strains; D013438:Sulfhydryl Compounds / Q000494:pharmacology*
D007527:Isoenzymes; D013438:Sulfhydryl Compounds; C042661:S-(propyl)glutathione; D007490:Iproniazid; D005982:Glutathione Transferase; D005978:Glutathione