title: Glutathione S-transferase conjugation of organophosphorus pesticides yields S-phospho-, S-aryl-, and S-alkylglutathione derivatives.
abstract: Pesticide detoxification is a central feature of selective toxicity and safety evaluation. Two of the principal enzymes involved are GSH S-transferases (GSTs) and cytochrome P450s acting alone and together. More than 100 pesticides are organophosphorus (OP) compounds, but with few exceptions, their GSH conjugates have not been directly observed in vitro or in vivo. The major insecticides chlorpyrifos (CP) and diazinon are of particular interest as multifunctional substrates with diverse metabolites, while ClP(S)(OEt) 2 and the cotton defoliant tribufos are possible precursors of phosphorylated GSH conjugates. Formation of GSH conjugates by GST with GSH was studied in vitro with and without metabolic activation by human liver microsomes or P450 3A4 with NADPH. Metabolites were analyzed by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). Five GSH conjugates were identified from CP and chlorpyrifos oxon (CPO), i.e., GSCP and GSCPO in which the 6-chloro substituent of CP and CPO, respectively, is displaced by GSH; S-(3,5,6-trichloropyridin-2-yl)glutathione; S-(3,5-dichloro-6-hydroxypyridin-2-yl)glutathione; and S-ethylglutathione. GST of a human liver microsomal preparation but not P450 3A4 with GSH metabolized CP to GSCP. With GST and GSH, diazinon and diazoxon gave S-(2-isopropyl-4-methylpyrimidin-6-yl)glutathione and ClP(S)(OEt) 2 yielded GSP(S)(OEt) 2. With microsomes, NADPH, GST, and GSH tribufos gave GSP(O)(SBu) 2. The liver of intraperitoneally treated mice contained GSCP from CP, GSP(S)(OEt) 2 from ClP(S)(OEt) 2, and GSP(O)(SBu) 2 from tribufos. GSP(S)(OEt) 2 and GSP(O)(SBu) 2 are the first S-phosphoglutathione metabolites observed in vitro and in vivo directly by LC-ESI-MS. Nine other OP pesticides gave only O-dealkylation in the GST/GSH system. GST-catalyzed metabolism joins P450s and hydrolases as important contributors to OP detoxification.
authors.affiliation: Environmental Chemistry and Toxicology Laboratory, Department of Environmental Science, Policy and Management, University of California, Berkeley, CA 94720-3112, USA.
authors.forename: Kazutoshi
authors.initials: K
authors.lastname: Fujioka
authors.forename: John E
authors.initials: JE
authors.lastname: Casida
mesh_terms: D000818:Animals; D004390:Chlorpyrifos / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity; D002853:Chromatography, Liquid; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000378:metabolism*; D003678:Defoliants, Chemical / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity; D003976:Diazinon / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism*; D005982:Glutathione Transferase / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006867:Hydrolases / Q000378:metabolism; D008658:Inactivation, Metabolic; D008099:Liver / Q000201:enzymology; D051379:Mice; D008861:Microsomes / Q000201:enzymology; D009249:NADP / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D010575:Pesticides / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D021241:Spectrometry, Mass, Electrospray Ionization; D013379:Substrate Specificity; D013997:Time Factors
chemical_list: D003678:Defoliants, Chemical; D009943:Organophosphorus Compounds; D010575:Pesticides; D009249:NADP; D003577:Cytochrome P-450 Enzyme System; D005982:Glutathione Transferase; D006867:Hydrolases; D005978:Glutathione; D004390:Chlorpyrifos; D003976:Diazinon