GSTP1-1 stereospecifically catalyzes glutathione conjugation of ethacrynic acid
article
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.
Topics
EA, ethacrynic acidEthacrynic acidGlutathione S-transferaseStereoselectivityEtacrynic acidGlutathioneGlutathione transferaseMultidrug resistance proteinChemical reactionConjugateControlled studyDiastereoisomerDrug conjugationEnzyme mechanismGlutathione metabolismHuman cellMelanoma cellNuclear magnetic resonance spectroscopyStereochemistryAmino Acid SubstitutionEthacrynic AcidGlutathioneGlutathione TransferaseHumansIsoenzymesMelanomaPoint MutationStereoisomerismSubstrate SpecificityTumor Cells, Cultured
TNO Identifier
234853
ISSN
00145793
Source
FEBS Letters, 441(1), pp. 153-157.
Pages
153-157