Greaves Jennifer, Tomkinson Nicholas C O
Faculty of Health and Life Sciences, Centre for Sport, Exercise and Life Sciences, Coventry University, Coventry, UK.
WestCHEM, Department of Pure and Applied Chemistry, University of Strathclyde, Glasgow, UK.
Methods Mol Biol. 2019;2009:13-33. doi: 10.1007/978-1-4939-9532-5_2.
The use of synthetically synthesized azide and alkyne fatty acid analogs coupled with bioorthogonal Cu(I)-catalyzed Huisgen 1,3-dipolar cycloaddition reaction-based detection methods to study protein S-acylation reactions has replaced the traditional method of using in vivo metabolic radiolabeling with tritiated palmitic acid and has greatly facilitated our understanding of this essential cellular process. Here, we describe the chemical synthesis of myristic (C:14), palmitic (C16:0), and stearic (C18:0) acid-azide probes and detail how they may be utilized as chemical reporters for the analysis of S-acylation of exogenously expressed proteins in cells.
使用化学合成的叠氮化物和炔烃脂肪酸类似物，结合基于生物正交铜（I）催化的胡伊斯根1,3-偶极环加成反应的检测方法来研究蛋白质S-酰化反应，已取代了使用氚标记的棕榈酸进行体内代谢放射性标记的传统方法，并极大地促进了我们对这一重要细胞过程的理解。在此，我们描述了肉豆蔻酸（C:14）、棕榈酸（C16:0）和硬脂酸（C18:0）-叠氮化物探针的化学合成，并详细介绍了如何将它们用作化学报告分子，以分析细胞中外源表达蛋白质的S-酰化作用。
