The intrinsic affinity between E2 and the Cys domain of E1 in ubiquitin-like modifications.
The intrinsic affinity between E2 and the Cys domain of E1 in ubiquitin-like modifications.
复制标题
DOI：
10.1016/j.molcel.2007.05.023
复制
发表时间：
2007-07
期刊：
影响因子：
16
通讯作者：
Jianghai Wang;Weidong Hu;S. Cai;Brian M. Lee;Jing Song;Yuan Chen
中科院分区：
文献类型：
--
作者：
Jianghai Wang;Weidong Hu;S. Cai;Brian M. Lee;Jing Song;Yuan Chen
类似泛素的修饰通过类似的生化机制进行，几乎调控细胞功能的方方面面。尽管最近在表征它们的酶学方面取得了进展，但我们对这些修饰的动态过程的了解仍然不完整。在这项研究中，我们揭示了相扑E2和相扑E1的半胱氨酸结构域之间的内在亲和力。核磁共振研究与顺磁自旋标记相结合表明，这种相互作用是由E1和E2上先前未知的界面介导的，并使两种酶的两个催化半胱氨酸残基非常接近。定点突变和酶分析表明，这种相互作用对相扑从E1转移到E2是至关重要的。本研究结果提示，在类泛素修饰中，E2与E1的半胱氨酸结构域的相互作用参与了引导E2‘S易位到E1’S酶活性部位的过程。
Ubiquitin-like modifications, which are carried out by similar biochemical mechanisms, regulate nearly every aspect of cellular function. Despite the recent advancements in characterizing their enzymology, our knowledge about the dynamic processes of these modifications is still fragmentary. In this study, we have uncovered an intrinsic affinity between the SUMO E2 and the Cys domain of SUMO E1. NMR studies in combination with paramagnetic spin labeling demonstrate that this interaction is mediated by previously unknown interfaces on both E1 and E2 and places the two catalytic Cys residues of the two enzymes in close proximity. Site-directed mutagenesis and enzymatic assays indicate that the interaction is fundamentally important for the transfer of SUMO from E1 to E2. Results from this study suggest that the interaction between E2 and the Cys domain of E1 participates in guiding the E2's translocation to E1's enzymatic active site in ubiquitin-like modifications.