Abrami Laurence, Denhardt-Eriksson Robin A, Hatzimanikatis Vassily, van der Goot F Gisou
Global Health Institute, School of Life Sciences, EPFL, Lausanne, Switzerland.
Global Health Institute, Laboratory of Computational Systems Biotechnology, EPFL, Lausanne, Switzerland.
Methods Mol Biol. 2019;2009:111-127. doi: 10.1007/978-1-4939-9532-5_9.
Proteins can be radiolabeled either during synthesis, typically using S-cysteine/methionine (S-Cys/Met), or after synthesis, by adding a radiolabeled posttranslational modification. Here we describe how protein S-palmitoylation, and its dynamics, can be monitored by H-palmitate labeling and how the importance of S-palmitoylation in protein biogenesis and turnover can be investigated using S-Cys/Met pulse-chase metabolic labeling. Proteins frequently have multiple palmitoylation sites. The importance thereof on the design and interpretation of metabolic labeling experiments is discussed.
蛋白质可以在合成过程中进行放射性标记，通常使用S-半胱氨酸/甲硫氨酸（S-Cys/Met），也可以在合成后通过添加放射性标记的翻译后修饰进行标记。在这里，我们描述了如何通过H-棕榈酸盐标记监测蛋白质S-棕榈酰化及其动态变化，以及如何使用S-Cys/Met脉冲追踪代谢标记研究S-棕榈酰化在蛋白质生物合成和周转中的重要性。蛋白质通常有多个棕榈酰化位点。本文讨论了其在代谢标记实验设计和解释中的重要性。
