title: GSTP1-1 stereospecifically catalyzes glutathione conjugation of ethacrynic acid.
abstract: Using 1H NMR two diastereoisomers of the ethacrynic acid glutathione conjugate (EASG) as well as ethacrynic acid (EA) could be distinguished and quantified individually. Chemically prepared EASG consists of equal amounts of both diastereoisomers. GSTP1-1 stereospecifically catalyzes formation of one of the diastereoisomers (A). The GSTP1-1 mutant C47S and GSTA1-1 preferentially form the same diastereoisomer of EASG as GSTP1-1. Glutathione conjugation of EA by GSTA1-2 and GSTA2-2 is not stereoselective. When human melanoma cells, expressing GSTP1-1, were exposed to ethacrynic acid, diastereoisomer A was the principal conjugate formed, indicating that even at physiological pH the enzyme catalyzed reaction dominates over the chemical conjugation.
authors.affiliation: Department of Food Technology and Nutritional Sciences, Wageningen Agricultural University, The Netherlands. marlou.vaniersel@algemeen.tox.wau.nl
authors.forename: M L
authors.initials: ML
authors.lastname: van Iersel
authors.forename: M M
authors.initials: MM
authors.lastname: van Lipzig
authors.forename: I M
authors.initials: IM
authors.lastname: Rietjens
authors.forename: J
authors.initials: J
authors.lastname: Vervoort
authors.forename: P J
authors.initials: PJ
authors.lastname: van Bladeren
mesh_terms: D019943:Amino Acid Substitution; D004976:Ethacrynic Acid / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism*; D005982:Glutathione Transferase / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism*; D008545:Melanoma / Q000201:enzymology; D017354:Point Mutation; D013237:Stereoisomerism; D013379:Substrate Specificity; D014407:Tumor Cells, Cultured
chemical_list: D007527:Isoenzymes; D005982:Glutathione Transferase; D005978:Glutathione; D004976:Ethacrynic Acid