Detection and characterization of SUMO protease activity using a sensitive enzyme-based reporter assay.
Detection and characterization of SUMO protease activity using a sensitive enzyme-based reporter assay.
复制标题
使用灵敏的基于酶的报告基因测定法检测和表征 SUMO 蛋白酶活性。
DOI：
10.1007/978-1-59745-566-4_18
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Nicholson,Benjamin
中科院分区：
文献类型：
--
作者：
Leach,CraigA;Tian,Xufan;Mattern,MichaelR;Nicholson,Benjamin
In this chapter we describe a novel, sensitive, homogenous high throughput reporter-basedin vitroassay for SUMO protease activity developed by Progenra, Inc. A reporter construct was created by fusing His6-tagged small ubiquitin-like modifi er (SUMO) to the amino terminus of the reporter enzyme phospholipase A2(PLA2). Following cleavage by a member of the sentrin specifi c proteases (SENPs), free PLA2is able to turn over its substrate, resulting in the release of a fl uorescent product which is readily quantifi able using a fl uorimeter or a fl uorescence plate reader. The utility of this SUMO-CHOP-Reporter assay platform is demonstrated by its ability to determine Kmvalues and to characterize inhibitors of SUMO proteases.
