Monitoring the Glutathione Redox Reaction in Living Human Cells by Combining Metabolic Labeling with Heteronuclear NMR |
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| Authors: | Xing Jin Soeun Kang Prof Shinya Tanaka Prof Sunghyouk Park |
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| Affiliation: | 1. College of Pharmacy, Natural Product Research Institute, Seoul National University, Seoul, Korea;2. Department of Pathology, Laboratory of Cancer Research, Hokkaido University Graduate School of Medicine, Sapporo, Japan |
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| Abstract: | The glutathione (GSH) redox reaction is critical for defense against cellular reactive oxygen species (ROS). However, direct and real‐time monitoring of this reaction in living mammalian cells has been hindered by the lack of a facile method. Herein, we describe a new approach that exploits the GSH biosynthetic pathway and heteronuclear NMR. U‐13C]‐labeled cysteine was incorporated into GSH in U87 glioblastoma cells, and the oxidation of GSH to GSSG by a ROS‐producing agent could be monitored in living cells. Further application of the approach to cells resistant to temozolomide (TMZ), an anti‐glioblastoma drug, suggested a possible new resistance mechanism involving neutralization of ROS. This result was corroborated by the observation of up‐regulation of glutathione peroxidase 3 (GPx3). This new approach could be easily applied to redox‐dependent signaling pathways and drug resistance involving ROS. |
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| Keywords: | cancer drug resistance glutathione in-cell NMR redox chemistry |
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