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Total Chemical Synthesis of SUMO and SUMO‐Based Probes for Profiling the Activity of SUMO‐Specific Proteases
Author(s) -
Mulder Monique P. C.,
Merkx Remco,
Witting Katharina F.,
Hameed Dharjath S.,
El Atmioui Dris,
Lelieveld Lindsey,
Liebelt Frauke,
Neefjes Jacques,
Berlin Ilana,
Vertegaal Alfred C. O.,
Ovaa Huib
Publication year - 2018
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201803483
Subject(s) - proteases , sumo protein , protease , computational biology , biochemistry , chemistry , sumo enzymes , biology , enzyme , microbiology and biotechnology , ubiquitin , gene
Abstract SUMO is a post‐translational modifier critical for cell cycle progression and genome stability that plays a role in tumorigenesis, thus rendering SUMO‐specific enzymes potential pharmacological targets. However, the systematic generation of tools for the activity profiling of SUMO‐specific enzymes has proven challenging. We developed a diversifiable synthetic platform for SUMO‐based probes by using a direct linear synthesis method, which permits N‐ and C‐terminal labelling to incorporate dyes and reactive warheads, respectively. In this manner, activity‐based probes (ABPs) for SUMO‐1, SUMO‐2, and SUMO‐3‐specific proteases were generated and validated in cells using gel‐based assays and confocal microscopy. We further expanded our toolbox with the synthesis of a K11‐linked diSUMO‐2 probe to study the proteolytic cleavage of SUMO chains. Together, these ABPs demonstrate the versatility and specificity of our synthetic SUMO platform for in vitro and in vivo characterization of the SUMO protease family.