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  • Title: Detection and characterization of SUMO protease activity using a sensitive enzyme-based reporter assay.
    Author: Leach CA, Tian X, Mattern MR, Nicholson B.
    Journal: Methods Mol Biol; 2009; 497():269-81. PubMed ID: 19107424.
    Abstract:
    In this chapter we describe a novel, sensitive, homogenous high throughput reporter-based in vitro assay for SUMO protease activity developed by Progenra, Inc. A reporter construct was created by fusing His(6)-tagged small ubiquitin-like modifier (SUMO) to the amino terminus of the reporter enzyme phospholipase A(2) (PLA(2)). Following cleavage by a member of the sentrin specific proteases (SENPs), free PLA(2) is able to turn over its substrate, resulting in the release of a fluorescent product which is readily quantifiable using a fluorimeter or a fluorescence plate reader. The utility of this SUMO-CHOP-Reporter assay platform is demonstrated by its ability to determine K(m) values and to characterize inhibitors of SUMO proteases.
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