Hemsley Piers A
Division of Plant Sciences, College of Life Sciences, University of Dundee, Dundee, UK.
Methods Mol Biol. 2013;1043:141-6. doi: 10.1007/978-1-62703-532-3_15.
S-acylation is increasingly being recognized as an important posttranslational modification of proteins controlling activity, subcellular localization, microdomain residence, and stability. Heterotrimeric G-proteins and GPCRs are particularly well studied S-acylated proteins, and fast, cheap, reliable methods are required for the analysis of S-acylation states of these proteins. Various approaches have been developed to study S-acylation, but they are time consuming, expensive, frequently require radiolabels and generally only suitable for cell culture, making them impractical for work in plant systems. Here a rapid and inexpensive method is described for the analysis of the S-acylation state of AGG2 that can be performed on any cell or tissue sample using standard laboratory equipment and methods. This method is also applicable to any protein that can be detected by western blotting.
S-酰化越来越被认为是一种重要的蛋白质翻译后修饰，它控制着蛋白质的活性、亚细胞定位、微结构域驻留和稳定性。异源三聚体G蛋白和G蛋白偶联受体（GPCR）是研究得特别透彻的S-酰化蛋白，因此需要快速、廉价且可靠的方法来分析这些蛋白的S-酰化状态。已经开发出各种方法来研究S-酰化，但它们耗时、昂贵，经常需要放射性标记，并且通常仅适用于细胞培养，这使得它们在植物系统中无法实际应用。本文描述了一种快速且廉价的方法，用于分析AGG2的S-酰化状态，该方法可使用标准实验室设备和方法在任何细胞或组织样本上进行。此方法也适用于任何可通过蛋白质免疫印迹法检测到的蛋白质。
