Chen Qian, Yang Xiaoyuan, Xie Qi
State Key Laboratory of Plant Genomics, National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, No. 1 West Beichen Road, Beijing, 100101, P. R. China.
Graduate University of Chinese Academy of Sciences, Beijing, 100049, P. R. China.
Methods Mol Biol. 2016;1450:3-10. doi: 10.1007/978-1-4939-3759-2_1.
Ubiquitination is an important posttranslational modification in eukaryotic organisms and plays a central role in many signaling pathways in plants. Most ubiquitination typically occurs on substrate lysine residues, forming a covalent isopeptide bond. Some recent reports suggested ubiquitin can be attached to non-lysine sites such as serine/threonine, cysteine or the N-terminal methionine, via oxyester or thioester linkages, respectively. In the present protocol, we developed a convenient in vitro assay for investigating ubiquitination on Ser/Thr and Cys residues.
泛素化是真核生物中一种重要的翻译后修饰，在植物的许多信号通路中起着核心作用。大多数泛素化通常发生在底物赖氨酸残基上，形成共价异肽键。最近的一些报道表明，泛素可以分别通过氧酯键或硫酯键连接到丝氨酸/苏氨酸、半胱氨酸或N端甲硫氨酸等非赖氨酸位点上。在本实验方案中，我们开发了一种简便的体外测定方法，用于研究丝氨酸/苏氨酸和半胱氨酸残基上的泛素化。
