Yamano Hiroyuki, Trickey Michelle, Grimaldi Margaret, Kimata Yuu
Cell Cycle Control Laboratory, Marie Curie Research Institute, Surrey, UK.
Methods Mol Biol. 2009;545:287-300. doi: 10.1007/978-1-60327-993-2_18.
The anaphase-promoting complex/cyclosome (APC/C), a large (20S) multisubunit E3 ligase, has an essential role to ubiquitylate numerous substrates at specific times during mitosis and G1 phase as well as in meiosis. The deregulation of the APC/C causes cell death or genomic instability, which is a hallmark of cancers. Although 13 years have passed since its discovery, the molecular mechanisms of the APC/C-dependent ubiquitylation and proteolysis are still poorly understood. The development of in vitro systems enables the identification of new substrates and investigation of the molecular mechanisms by which the APC/C recognizes its substrates. This chapter describes in vitro assays reconstituted in Xenopus egg extracts.
后期促进复合物/细胞周期体（APC/C）是一种大型（20S）多亚基E3连接酶，在有丝分裂、G1期以及减数分裂的特定时间对众多底物进行泛素化起着至关重要的作用。APC/C失调会导致细胞死亡或基因组不稳定，而这是癌症的一个标志。尽管自其发现以来已过去13年，但对APC/C依赖性泛素化和蛋白水解的分子机制仍知之甚少。体外系统的发展使得能够鉴定新底物，并研究APC/C识别其底物的分子机制。本章描述了在非洲爪蟾卵提取物中重建的体外测定方法。
