RWD Domain as an E2 (Ubc9)-Interaction Module
RWD Domain as an E2 (Ubc9)-Interaction Module
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DOI：
10.1074/jbc.m115.644047
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发表时间：
2015-07-03
影响因子：
4.8
通讯作者：
Chen, Yuan
中科院分区：
文献类型：
--
作者：
Alontaga, Aileen Y.;Ambaye, Nigus D.;Chen, Yuan
RWD结构域是通过人类蛋白质组序列的生物信息学分析发现的高度保守的结构域;然而，其功能尚不清楚。泛素样修饰需要三种酶的催化，通常称为E1，E2和E3。我们解决了与RWD结构域复合的小泛素样修饰剂（SUMO）的E2的晶体结构，并使用溶液NMR分析确认了结构。Ubc 9上RWD的结合表面位于Ubc 9的N末端附近，已知其参与缀合机制中蛋白质的非共价结合，包括E1、SUMO和E3连接酶的结构域。NMR数据表明RWD结构域不与SUMO和E1结合。RWD与Ubc 9之间的相互作用具有32 +/- 4 μ M的Kd。与结构和结合亲和力一致，并且与先前的报道相反，RWD结构域和RWDD 3对全局SUMO化的影响最小。这里提出的结构和生物化学信息形成的基础上，进一步调查的功能RWD含蛋白质。
An RWD domain is a well conserved domain found through bioinformatic analysis of the human proteome sequence; however, its function has been unknown. Ubiquitin-like modifications require the catalysis of three enzymes generally known as E1, E2, and E3. We solved the crystal structure of the E2 for the small ubiquitin-like modifiers (SUMO) in complex with an RWD domain and confirmed the structure using solution NMR analysis. The binding surface of RWD on Ubc9 is located near the N terminus of Ubc9 that is known to be involved in non-covalent binding of the proteins in the conjugation machinery, including a domain of E1, SUMO, and an E3 ligase. NMR data indicate that the RWD domain does not bind to SUMO and E1. The interaction between RWD and Ubc9 has a K-d of 32 +/- 4 mu M. Consistent with the structure and binding affinity and in contrast to a previous report, the RWD domain and RWDD3 have minimal effects on global SUMOylation. The structural and biochemical information presented here forms the basis for further investigation of the functions of RWD-containing proteins.