Assay methods for small ubiquitin-like modifier (SUMO)-SUMO-interacting motif (SIM) interactions in vivo and in vitro using a split-luciferase complementation system
Assay methods for small ubiquitin-like modifier (SUMO)-SUMO-interacting motif (SIM) interactions in vivo and in vitro using a split-luciferase complementation system
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DOI：
10.1016/j.ab.2013.12.009
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发表时间：
2014-03-01
影响因子：
2.9
通讯作者：
Yoshida, Minoru
中科院分区：
文献类型：
--
作者：
Hirohama, Mikako;Voet, Arnout R. D.;Yoshida, Minoru
SUMO 化是一种翻译后过程，它将一个小的泛素样修饰剂 (SUMO) 共价连接到其靶蛋白上。 SUMO 化通过 SUMO 相互作用基序 (SIM) 识别 SUMO 来控制多个细胞过程。在本研究中，我们开发了利用分裂荧光素酶互补检测细胞中 SUMO 和 SIM 之间非共价相互作用的检测系统。我们应用该测定的一个版本来使用细菌表达系统检测体外 SUMO-SIM 相互作用。这些新颖的检测方法能够以高通量方式在体内或体外筛选 SUMO 依赖性蛋白质-蛋白质相互作用的抑制剂。 (C) 2013 Elsevier Inc. 保留所有权利。
SUMOylation is a posttranslational process that attaches a small ubiquitin-like modifier (SUMO) to its target proteins covalently. SUMOylation controls multiple cellular processes through the recognition of SUMO by a SUMO-interacting motif (SIM). In this study, we developed assay systems for detecting noncovalent interactions between SUMO and SIM in cells using split-luciferase complementation. We applied a version of this assay to the detection of in vitro SUMO-SIM interactions using a bacterial expression system. These novel assays enable screening of inhibitors of SUMO-dependent protein-protein interactions, either in vivo or in vitro, in a high-throughput manner. (C) 2013 Elsevier Inc. All rights reserved.