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The Biochemical journal1990; 271(1); 161-165; doi: 10.1042/bj2710161

Inhibition and recognition studies on the glutathione-binding site of equine liver glutathione S-transferase.

Abstract: Equine liver glutathione S-transferase has been shown to consist of two identical subunits of apparent Mr 25,500 and a pl of 8.9. Kinetic data at pH 6.5 with 1-chloro-2,4-dinitrobenzene as a substrate suggests a random rapid-equilibrium mechanism, which is supported by inhibition studies using glutathione analogues. S-(p-Bromobenzyl)glutathione and the corresponding N alpha-, CGlu- and CGly-substituted derivatives have been found, at pH 6.5, to be linear competitive inhibitors, with respect to GSH, of glutathione transferase. N-Acetylation of S-(p-bromobenzyl)glutathione decreases binding by 100-fold, whereas N-benzoylation and N-benzyloxycarbonylation abolish binding of the derivative to the enzyme. The latter effect has been attributed to a steric constraint in this region of the enzyme. Amidation of the glycine carboxy group of S-(p-bromobenzyl)glutathione decreases binding by 13-fold, whereas methylation decreases binding by 70-fold, indicating a steric constraint and a possible electrostatic interaction in this region of the enzyme. Amidation of both carboxy groups decreases binding significantly by 802-fold, which agrees with electrostatic interaction of the glutamic acid carboxy group with a group located on the enzyme.
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Publication Date: 1990-10-01 PubMed ID: 2222409PubMed Central: PMC1149527DOI: 10.1042/bj2710161Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research explores the interaction and inhibition of equine liver glutathione S-transferase with glutathione analogues, where different glutathione derivatives showed varied inhibitory effects.

Research Background

  • The study is centered around equine liver glutathione S-transferase (GST), an enzyme involved in the detoxification process in the body. This enzyme is made up of two identical subunits each with a molecular weight of 25,500 and an iso-electric point (pI) of 8.9.
  • Using 1-chloro-2,4-dinitrobenzene as a substrate, the researchers studied the mechanics of the enzyme at pH 6.5 and inferred a random rapid-equilibrium mechanism.

Inhibition Studies

  • The study worked with glutathione (GSH) analogues in inhibition studies to understand the mechanism of the enzyme. S-(p-Bromobenzyl)glutathione and its various modified alternatives were used as glutathione analogues.
  • At pH 6.5, these compounds were found to be linear competitive inhibitors for GST, meaning they compete with glutathione in binding to the enzyme.

Effects of Glutathione Derivative Modifications

  • The research found that changing specific molecular groups on the glutathione analogue effected its binding to GST. For example, N-acetylation resulted in a 100-fold decrease in binding affinity.
  • N-benzoylation and N-benzyloxycarbonylation completely eliminated binding to the enzyme, indicating that these modifications introduced a steric hinderance that prevented the molecule from fitting in the active site of the enzyme.
  • Amidation and methylation of the glycine carboxy group led to a 13-fold and 70-fold decrease in binding, respectively, implying both steric and possible electrostatic constraints at this part of the enzyme.
  • Amidating both carboxy groups led to a dramatic 802-fold decrease in binding, which suggests an important role of the electrostatic interaction of the glutamic acid carboxy group with the enzyme.

Conclusion

  • The examination of the reaction mechanism and the binding behavior of glutathione derivatives to equine liver GST could enhance our understanding of the blocking or inhibiting potential of certain compounds to GST.

Cite This Article

APA
D'Silva C. (1990). Inhibition and recognition studies on the glutathione-binding site of equine liver glutathione S-transferase. Biochem J, 271(1), 161-165. https://doi.org/10.1042/bj2710161

Publication

The Biochemical journal
ISSN: 0264-6021
NlmUniqueID: 2984726R
Country: England
Language: English
Volume: 271
Issue: 1
Pages: 161-165

Researcher Affiliations

D'Silva, C
  • Institute of Molecular and Biomolecular Electronics, University of Wales, Bangor, Gwynedd, U.K.

MeSH Terms

  • Animals
  • Binding Sites
  • Binding, Competitive
  • Dinitrochlorobenzene / metabolism
  • Glutathione / analogs & derivatives
  • Glutathione / chemistry
  • Glutathione / metabolism
  • Glutathione Transferase / antagonists & inhibitors
  • Glutathione Transferase / metabolism
  • Horses
  • Kinetics
  • Liver / enzymology
  • Molecular Weight
  • Substrate Specificity

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Citations

This article has been cited 4 times.
  1. Penketh PG, Patridge E, Shyam K, Baumann RP, Zhu R, Ishiguro K, Sartorelli AC. Influence of glutathione and glutathione S-transferases on DNA interstrand cross-link formation by 1,2-bis(methylsulfonyl)-1-(2-chloroethyl)hydrazine, the active anticancer moiety generated by laromustine. Chem Res Toxicol 2014 Aug 18;27(8):1440-9.
    doi: 10.1021/tx500197tpubmed: 25012050google scholar: lookup
  2. Gusson F, Carletti M, Albo AG, Dacasto M, Nebbia C. Comparison of hydrolytic and conjugative biotransformation pathways in horse, cattle, pig, broiler chick, rabbit and rat liver subcellullar fractions. Vet Res Commun 2006 Apr;30(3):271-83.
    doi: 10.1007/s11259-006-3247-ypubmed: 16437303google scholar: lookup
  3. D'Silva C. Synthesis of carboxy-residue-modified coenzyme derivatives as probes to the mechanism of glutathione enzymes. Biochem J 1990 Oct 1;271(1):167-9.
    doi: 10.1042/bj2710167pubmed: 2222410google scholar: lookup
  4. Polhuijs M, Lankhaar G, Mulder GJ. Relationship between glutathione content in liver and glutathione conjugation rate in the rat in vivo. Effect of buthionine sulphoximine pretreatment on conjugation of the two 2-bromoisovalerylurea enantiomers during intravenous infusion. Biochem J 1992 Jul 15;285 ( Pt 2)(Pt 2):401-4.
    doi: 10.1042/bj2850401pubmed: 1637334google scholar: lookup
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