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GSTP1-1 stereospecifically catalyzes glutathione conjugation of ethacrynic acid

Author: Iersel, M.L.P.S. van · Lipzig, M.M.H. van · Rietjens, I.M.C.M. · Vervoort, J. · Bladeren, P.J. van
Type:article
Date:1998
Institution: Centraal Instituut voor Voedingsonderzoek TNO
Source:FEBS Letters, 1, 441, 153-157
Identifier: 234853
doi: doi:10.1016/S0014-5793(98)01546-4
Keywords: EA, ethacrynic acid · Ethacrynic acid · Glutathione S-transferase · Stereoselectivity · Etacrynic acid · Glutathione · Glutathione transferase · Multidrug resistance protein · Chemical reaction · Conjugate · Controlled study · Diastereoisomer · Drug conjugation · Enzyme mechanism · Glutathione metabolism · Human cell · Melanoma cell · Nuclear magnetic resonance spectroscopy · Stereochemistry · Amino Acid Substitution · Ethacrynic Acid · Glutathione · Glutathione Transferase · Humans · Isoenzymes · Melanoma · Point Mutation · Stereoisomerism · Substrate Specificity · Tumor Cells, Cultured

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. Copyright (C) 1998 Federation of European Biochemical Societies.