1. Direct evidence for the covalent modification of glutathione-S-transferase P1-1 by electrophilic prostaglandins: Implications for enzyme inactivation and cell survival
- Author
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Javier Machín Gayarre, M. Isabel Avellano, Dolores Pérez-Sala, and Francisco J. Sánchez-Gómez
- Subjects
Cyclopentenone ,Cell Survival ,Molecular Sequence Data ,Biophysics ,Antineoplastic Agents ,Endogeny ,Cyclopentanes ,urologic and male genital diseases ,Biochemistry ,Cell Line ,Mice ,chemistry.chemical_compound ,Animals ,Humans ,Amino Acid Sequence ,neoplasms ,Molecular Biology ,Cyclopentenone prostaglandins ,chemistry.chemical_classification ,biology ,Prostaglandin D2 ,Glutathione ,Recombinant Proteins ,Enzyme Activation ,Glutathione S-transferase ,Enzyme ,Glutathione S-Transferase pi ,chemistry ,Covalent bond ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Biotinylation ,biology.protein ,Protein Binding - Abstract
Glutathione-S-transferases (GST) catalyze the conjugation of electrophilic compounds to glutathione, thus playing a key role in cell survival and tumor chemoresistance. Cyclopentenone prostaglandins (cyPG) are electrophilic eicosanoids that display potent antiproliferative properties, through multiple mechanisms not completely elucidated. Here we show that the cyPG 15-deoxy-Δ12,14-PGJ2 (15d-PGJ2) binds to GSTP1-1 covalently, as demonstrated by mass spectrometry and by the use of biotinylated 15d-PGJ2. Moreover, cyPG inactivate GSTP1-1 irreversibly. The presence of the cyclopentenone moiety is important for these effects. Covalent interactions also occur in cells, in which 15d-PGJ2 binds to endogenous GSTP1-1, irreversibly reduces GST free-thiol content and inhibits GST activity. Protein delivery of GSTP1-1 improves cell survival upon serum deprivation whereas 15d-PGJ2-treated GSTP1-1 displays a reduced protective effect. These results show the first evidence for the formation of stable adducts between cyPG and GSTP1-1 and may offer new perspectives for the development of irreversible GST inhibitors as anticancer agents.
- Published
- 2007