放射性代谢标记法同时检测磷酯酰肌醇脂。
Simultaneous Detection of Phosphoinositide Lipids by Radioactive Metabolic Labeling.
机构信息
Life Sciences Institute, University of Michigan, Ann Arbor, MI, USA.
Program in Cellular and Molecular Biology, University of Michigan, Ann Arbor, MI, USA.
出版信息
Methods Mol Biol. 2021;2251:1-17. doi: 10.1007/978-1-0716-1142-5_1.
Abstract
Phosphoinositide (PPI) lipids are a crucial class of low-abundance signaling molecules that regulate many processes within cells. Methods that enable simultaneous detection of all PPI lipid species provide a wholistic snapshot of the PPI profile of cells, which is critical for probing PPI biology. Here we describe a method for the simultaneous measurement of cellular PPI levels by metabolically labeling yeast or mammalian cells with myo-H-inositol, extracting radiolabeled glycerophosphoinositides, and separating lipid species on an anion exchange column via HPLC.
摘要
磷酸肌醇（PPI）脂质是一类重要的低丰度信号分子，它们调节细胞内的许多过程。能够同时检测所有 PPI 脂质种类的方法提供了细胞 PPI 谱的整体快照，这对于探测 PPI 生物学至关重要。在这里，我们描述了一种通过用肌-肌醇- 氚标记酵母或哺乳动物细胞、提取放射性标记的甘油磷酸肌醇和在阴离子交换柱上通过 HPLC 分离脂质种类来同时测量细胞内 PPI 水平的方法。
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