Highly sensitive assays for SUMOylation and small ubiquitin-like modifier-dependent protein-protein interactions.
Highly sensitive assays for SUMOylation and small ubiquitin-like modifier-dependent protein-protein interactions.
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DOI：
10.1016/j.ab.2007.11.024
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发表时间：
2008-04
影响因子：
2.9
通讯作者：
N. Rouleau;Jianghai Wang;Labrini Karras;E. Andrews;M. Bielefeld-Sévigny;Yuan Chen
中科院分区：
文献类型：
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作者：
N. Rouleau;Jianghai Wang;Labrini Karras;E. Andrews;M. Bielefeld-Sévigny;Yuan Chen
小泛素样蛋白（SUMO）是近年来发现的一类重要的翻译后修饰蛋白，参与调节蛋白质的功能和细胞内转运。在这项研究中，我们描述了两种基于化学发光的检测方法，一种用于SUMO化，另一种用于SUMO介导的蛋白质-蛋白质相互作用。这些测定可用于表征催化SUMO化的酶的活性和动力学，以及用于高通量筛选SUMO化和SUMO依赖性蛋白质-蛋白质相互作用的抑制剂。这些新的检测方法代表了迄今为止发表的最灵敏的泛素样系统的检测方法。类似的策略可以用于开发其他泛素样修饰系统的测定。
Small ubiquitin-like proteins (SUMO) are recently discovered post-translational modifiers that regulate protein functions and intracellular trafficking. In this study, we are describing two chemoluminescence-based assays, one for SUMOylation and another one for SUMO-mediated protein-protein interactions. These assays can be used to characterize the activity and kinetics of the enzymes that catalyze SUMOylation, and in high-throughput screening for inhibitors of SUMOylation and SUMO-dependent protein-protein interactions. These novel assays represent the most sensitive assays for ubiquitin-like systems published to date. Similar strategies can be used to develop assays for other ubiquitin-like modification systems.