Redox Metabolism Measurement in Mammalian Cells and Tissues by LC-MS
| Autor: | Alan Wong, Anna Warren, Naama Kanarek, Boryana Petrova, Andrew J. Culhane, Nuria Yulia Vital, Adam G. Maynard |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Endocrinology Diabetes and Metabolism Cell medicine.disease_cause Biochemistry Redox Microbiology Article 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine redox metabolite detection in mammalian cells medicine NADPH redox metabolites glutathione mass-spectrometry method Molecular Biology chemistry.chemical_classification Reactive oxygen species Glutathione Small molecule QR1-502 030104 developmental biology medicine.anatomical_structure chemistry NADH Signal transduction HILIC chromatography 030217 neurology & neurosurgery Function (biology) Oxidative stress |
| Zdroj: | Metabolites, Vol 11, Iss 313, p 313 (2021) Metabolites Volume 11 Issue 5 |
| ISSN: | 2218-1989 |
| Popis: | Cellular redox state is highly dynamic and delicately balanced between constant production of reactive oxygen species (ROS), and neutralization by endogenous antioxidants, such as glutathione. Physiologic ROS levels can function as signal transduction messengers, while high levels of ROS can react with and damage various molecules eliciting cellular toxicity. The redox state is reflective of the cell’s metabolic status and can inform on regulated cell-state transitions or various pathologies including aging and cancer. Therefore, methods that enable reliable, quantitative readout of the cellular redox state are imperative for scientists from multiple fields. Liquid-chromatography mass-spectrometry (LC-MS) based methods to detect small molecules that reflect the redox balance in the cell such as glutathione, NADH, and NADPH, have been developed and applied successfully, but because redox metabolites are very labile, these methods are not easily standardized or consolidated. Here, we report a robust LC-MS method for the simultaneous detection of several redox-reactive metabolites that is compatible with parallel global metabolic profiling in mammalian cells. We performed a comprehensive comparison between three commercial hydrophilic interaction chromatography (HILIC) columns, and we describe the application of our method in mammalian cells and tissues. The presented method is easily applicable and will enable the study of ROS function and oxidative stress in mammalian cells by researchers from various fields. |
| Databáze: | OpenAIRE |
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