1. γ-Glutamyltranspeptidase-Dependent Metabolism of 4-Hydroxynonenal–Glutathione Conjugate
- Author
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Claude Marson, Haline Bodaud, Maria Wellman, Gérard Siest, Régine Herber, Pierre Leroy, Robert Wennig, Niculina Mitrea, and Milica Enoiu
- Subjects
Time Factors ,Cell Survival ,Metabolite ,Biophysics ,Tetrazolium Salts ,Cysteine Proteinase Inhibitors ,digestive system ,Biochemistry ,Mass Spectrometry ,Cell Line ,4-Hydroxynonenal ,Lipid peroxidation ,chemistry.chemical_compound ,Cricetinae ,Nonenal ,Animals ,Humans ,Cytotoxic T cell ,Coloring Agents ,Molecular Biology ,Chromatography, High Pressure Liquid ,Aldehydes ,Dose-Response Relationship, Drug ,Chemistry ,gamma-Glutamyltransferase ,Metabolism ,Glutathione ,Fibroblasts ,respiratory system ,digestive system diseases ,Kinetics ,Thiazoles ,Models, Chemical ,Spectrophotometry ,Cell culture ,Protein Binding - Abstract
A major pathway for detoxification of the highly reactive lipid peroxidation product, 4-hydroxy-2,3- trans -nonenal (HNE) is through the conjugation with glutathione (GSH). We have studied the metabolism of GS-HNE conjugate by the enzyme gamma-glutamyltranspeptidase (GGT) using its purified form, as well as a GGT-overexpressing fibroblast cell line (V79 GGT). Using mass spectrometry analysis we identified for the first time cysteinylglycine-HNE (CysGly-HNE) as the GGT metabolite of GS-HNE. Furthermore, the GGT-dependent metabolism of GS-HNE in the V79 GGT cell line was associated with a considerable increase of cytotoxicity as compared to a control cell line which does not express GGT (V79 Cl). The cytotoxic effect was dose- and time-dependent (100% cellular death at 200 μM GS-HNE after 24 h incubation) in V79 GGT cells, whereas no decrease of viability was observed in V79 Cl cells. A similar cytotoxic effect was obtained when cells were incubated directly with CysGly-HNE, demonstrating that this GGT-dependent metabolite unlike GS-HNE, exhibits cytotoxic properties.
- Published
- 2002
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