The Three-Dimensional Structure of the Human Pi Class Glutathione Transferase P1-1 in Complex with the Inhibitor Ethacrynic Acid and Its Glutathione Conjugate<sup>,</sup>

Oakley, Aaron J.; Rossjohn, Jamie; Bello, Mario Lo; Caccuri, Anna Maria; Federici, Giorgio; Parker, Michael W.

doi:10.1021/bi962316i

Cited by 122 publications

(121 citation statements)

References 39 publications

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“…The crystal structures of both hGSTA1-1 (Cameron et al 1995) and hGSTP1-1 (Oakley et al 1997c) in complex with the diuretic drug EA and its GSH-conjugate (GS-EA) have been determined (Figure 3b,c,d ,e). These studies were among the first to demonstrate that the H-sites of the pi-and alpha-class GSTs bind similar compounds distinctly.…”

Section: Gsts In Detoxificationmentioning

confidence: 99%

Glutathione transferases: a structural perspective

Oakley

2011

Drug Metabolism Reviews

316

240

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The glutathione transferases (GSTs) are one of the most important families of detoxifying enzymes in nature. The classic activity of the GSTs is conjugation of compounds with electrophilic centers to the tripeptide glutathione (GSH), but many other activities are now associated with GSTs, including steroid and leukotriene biosynthesis, peroxide degradation, double-bond cis-trans isomerization, dehydroascorbate reduction, Michael addition, and noncatalytic "ligandin" activity (ligand binding and transport). Since the first GST structure was determined in 1991, there has been an explosion in structural data across GSTs of all three families: the cytosolic GSTs, the mitochondrial GSTs, and the membrane-associated proteins in eicosanoid and glutathione metabolism (MAPEG family). In this review, the major insights into GST structure and function will be discussed. AbstractThe glutathione transferases (GSTs) are one of the most important families of detoxifying enzymes in nature. The classic activity of the GSTs is conjugation of compounds with electrophilic centers to the tripeptide glutathione (GSH), but many other activities are now associated with GSTs, including steroid and leukotriene biosynthesis, peroxide degradation, double-bond cis-trans isomerization, dehydroascorbate reduction, Michael addition, and non-catalytic "ligandin" activity (ligand binding and transport). Since the first GST structure was determined in 1991, there has been an explosion in structural data across GSTs of all three families: the cytosolic GSTs, the mitochondrial GSTs and the membrane-associated proteins in eicosanoid and glutathione metabolism (MAPEG family). In this review, the major insights into GST structure and function will be discussed.

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Section: Gsts In Detoxificationmentioning

confidence: 99%

Glutathione transferases: a structural perspective

Oakley

2011

Drug Metabolism Reviews

316

240

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“…The three-dimensional structure of the human GST P1-1, in complex with the inhibitor ETA, implicated that the inhibitor sits in a hydrophobic pocket lined with the side chains of Tyr7, Phe8, Pro9, and Val10, while the aliphatic portions of Arg13, Val35, Ile104, and Tyr108, and the carboxylic acid moiety of the inhibitor forms a hydrogen bond to the Nε atom of Arg13. 29 Moreover, two electrostatic interactions were observed in hGST P1-1[V104] with a GSH conjugate of (+)-anti-7,8-dihydroxy-9,10-oxy-7,8,9,10-tetrahydrobenzo [α]pyrene (GSBpd), including a hydrogen bond between the hydroxyl group of Tyr7 and the sulfur atom of GSH and a direct hydrogen bond between Arg13 and the hydroxyl group of the Bpd moiety of GSBpd, demonstrating participation of Arg13 in catalysis. 13 Similarly, Bjőrnestedt et al reported mutations of the Arg15 residue (the counterpart of Arg13 in GST P1-1) in human class Alpha GST A1-1.…”

Section: S-hexylgshmentioning

confidence: 99%

Contribution of Arginine 13 to the Catalytic Activity of Human Class Pi Glutathione Transferase P1-1

Kong¹,

Jo²,

Do³

et al. 2010

Bulletin of the Korean Chemical Society

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Arg13 is a conserved active-site residue in all known Pi class glutathione S-transferases (GSTs) and in most Alpha class GSTs. To evaluate its contribution to substrate binding and catalysis of this residue, three mutants (R13A, R13K, and R13L) were expressed in Escherichia coli and purified by GSH affinity chromatography. The substitutions of Arg13 significantly affected GSH-conjugation activity, while scarcely affecting glutathione peroxidase or steroid isomerase activities. Mutation of Arg13 into Ala largely reduced the GSH-conjugation activity by approximately 85 -95%, whereas substitutions by Lys and Leu barely affected activity. These results suggest that, in the GSH-conjugation activity of hGST P1-1, the contribution of Arg13 toward catalytic activity is highly dependent on substrate specificities and the size of the side chain at position 13. From the kinetic parameters, introduction of larger side chains at position 13 results in stronger affinity (Leu > Lys, Arg > Ala) towards GSH. The substitutions of Arg13 with alanine and leucine significantly affected kcat, whereas substitution with Lys was similar to that of the wild type, indicating the significance of a positively charged residue at position 13. From the plots of log (kcat/Km CDNB ) against pH, the pKa values of the thiol group of GSH bound in R13A, R13K, and R13L were estimated to be 1.8, 1.4, and 1.8 pK units higher than the pKa value of the wildtype enzyme, demonstrating the contribution of the Arg13 guanidinium group to the electrostatic field in the active site. From these results, we suggest that contribution of Arg13 in substrate binding is highly dependent on the nature of the electrophilic substrates, while in the catalytic mechanism, it stabilizes the GSH thiolate through hydrogen bonding.

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“…The observation that the diagnostic GST substrate CDNB does not protect against inactivation even though KPF-SCoA is itself a GST substrate can be explained in view of some previous works. For instance, it has been reported that ethacrynic acid, both a GST inhibitor and a GST substrate, can bind to human GST P1-1 in a nonproductive and a productive mode, which suggests the existence of multiple binding sites and binding modes for GST ligands (Oakley et al, 1997). The KPF-SCoA binding mode and binding stoichiometry may thus be distinct from those of CDNB.…”

Section: Interaction Of Ketoprofen Acylated Metabolites With Gstmentioning

confidence: 99%

Interaction of the Electrophilic Ketoprofenyl-Glucuronide and Ketoprofenyl-Coenzyme A Conjugates with Cytosolic GlutathioneS-Transferases

Osbild¹,

Bour²,

Maunit³

et al. 2007

Drug Metab Dispos

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ABSTRACT:Carboxylic acid-containing drugs are metabolized mainly through the formation of glucuronide and coenzyme A esters. These conjugates have been suspected to be responsible for the toxicity of several nonsteroidal anti-inflammatory drugs because of the reactivity of the electrophilic ester bond. In the present study we investigated the reactivity of ketoprofenyl-acylglucuronide (KPF-OG) and ketoprofenyl-acyl-coenzyme A (KPF-SCoA) toward cytosolic rat liver glutathione S-transferases (GST). We observed that KPFSCoA, but not KPF-OG inhibited the conjugation of 1-chloro-2,4-dinitrobenzene and 4-nitroquinoline N-oxide catalyzed by both purified cytosolic rat liver GST and GST from FAO and H5-6 rat hepatoma cell lines. Photoaffinity labeling with KPF-SCoA suggested that the binding of this metabolite may overlap the binding site of 4-methylumbelliferone sulfate. Furthermore, high-performance liquid chromatography and mass spectrometry analysis showed that both hydrolysis and transacylation reactions were observed in the presence of GST and glutathione. The formation of ketoprofenyl-S-acyl-glutathione could be kinetically characterized (apparent K m ‫؍‬ 196.0 ؎ 70.6 M). It is concluded that KPF-SCoA is both a GST inhibitor and a substrate of a GST-dependent transacylation reaction. The reactivity and inhibitory potency of thioester CoA derivatives toward GST may have potential implications on the reported in vivo toxicity of some carboxylic acid-containing drugs.

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The Three-Dimensional Structure of the Human Pi Class Glutathione Transferase P1-1 in Complex with the Inhibitor Ethacrynic Acid and Its Glutathione Conjugate^,

Cited by 122 publications

References 39 publications

Glutathione transferases: a structural perspective

Glutathione transferases: a structural perspective

Contribution of Arginine 13 to the Catalytic Activity of Human Class Pi Glutathione Transferase P1-1

Interaction of the Electrophilic Ketoprofenyl-Glucuronide and Ketoprofenyl-Coenzyme A Conjugates with Cytosolic GlutathioneS-Transferases

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The Three-Dimensional Structure of the Human Pi Class Glutathione Transferase P1-1 in Complex with the Inhibitor Ethacrynic Acid and Its Glutathione Conjugate,

Cited by 122 publications

References 39 publications

Glutathione transferases: a structural perspective

Glutathione transferases: a structural perspective

Contribution of Arginine 13 to the Catalytic Activity of Human Class Pi Glutathione Transferase P1-1

Interaction of the Electrophilic Ketoprofenyl-Glucuronide and Ketoprofenyl-Coenzyme A Conjugates with Cytosolic GlutathioneS-Transferases

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The Three-Dimensional Structure of the Human Pi Class Glutathione Transferase P1-1 in Complex with the Inhibitor Ethacrynic Acid and Its Glutathione Conjugate^,