Department of Chemistry, University of Washington, Seattle, WA, USA.
Methods Mol Biol. 2020;2133:293-312. doi: 10.1007/978-1-0716-0434-2_14.
The posttranslational modification of cellular proteins by ubiquitin (Ub), called ubiquitylation, is indispensable for the normal growth and development of eukaryotic organisms. In order to conduct studies that elucidate the precise mechanistic roles for Ub, access to site-specifically and homogenously ubiquitylated proteins and peptides is critical. However, the low abundance, heterogeneity, and dynamic nature of protein ubiquitylation are significant limitations toward such studies. Here we provide a facile expressed protein ligation method that does not require specialized apparatus and permits the rapid semisynthesis of ubiquitylated peptides by using the atom-efficient ligation auxiliary 2-aminooxyethanethiol.
细胞蛋白的泛素（Ub）翻译后修饰，称为泛素化，对真核生物的正常生长和发育是必不可少的。为了进行阐明 Ub 精确的机械作用的研究，获得特异性和均一地泛素化蛋白质和肽至关重要。然而，蛋白质泛素化的低丰度、异质性和动态性质是这些研究的重大限制。在这里，我们提供了一种简便的表达蛋白连接方法，该方法不需要专门的设备，并允许使用高效原子连接辅助物 2-氨基乙硫醇快速半合成泛素化肽。
