Quantification and Tracing of Stable Isotope into Cysteine and Glutathione.
Quantification and Tracing of Stable Isotope into Cysteine and Glutathione.
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半胱氨酸和谷胱甘肽稳定同位素的定量和追踪。
DOI：
10.1007/978-1-0716-3247-5_5
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
DeNicola,GinaM
中科院分区：
文献类型：
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作者：
Kang,YunPyo;DeNicola,GinaM
生物样品中代谢扰动的分析对于理解代谢性疾病的机制至关重要。在这里，我们描述了一个协议的定量稳定同位素标记的代谢物示踪培养细胞中的半胱氨酸代谢。该方案依赖于提取方案来衍生游离硫醇以防止氧化。此外，定量跟踪丝氨酸进入多个途径，包括谷胱甘肽合成途径，允许询问半胱氨酸和谷胱甘肽的合成。该方案提供了一个灵活的框架，可适用于询问许多代谢物和感兴趣的途径。
The analysis of metabolic perturbation in biological samples is crucial to understand mechanisms of metabolic diseases. Here, we describe a protocol for quantitative stable isotope-labeled metabolite tracing of cysteine metabolism in cultured cells. This protocol relies on an extraction protocol to derivatize free thiols to prevent oxidation. In addition, the quantitative tracing of serine into multiple pathways, including the glutathione synthesis pathway, allows for the interrogation of cysteine and glutathione synthesis. This protocol provides a flexible framework that can be adapted to interrogate many metabolites and pathways of interest.