Release of Enzymatically Active Deubiquitinating Enzymes Upon Reversible Capture by Disulfide Ubiquitin Reagents

Overview

Journal Angew Chem Int Ed Engl

Specialty Chemistry

Date 2017 Aug 26

PMID 28841265

Citations 13

Authors

Annemieke de Jong

Katharina Witting

Raymond Kooij

Dennis Flierman

Huib Ovaa

Affiliations

Soon will be listed here.

Abstract

Deubiquitinating enzymes (DUBs) catalyze the cleavage of ubiquitin from target proteins. Ubiquitin is post-translationally attached to proteins and serves as an important regulatory signal for key cellular processes. In this study, novel activity-based probes to study DUBs were synthesized that comprise a ubiquitin moiety and a novel disulfide warhead at the C-terminus. These reagents can bind DUBs covalently by forming a disulfide bridge between the active-site cysteine residue and the ubiquitin-based probe. As disulfide bridges can be broken by the addition of a reducing agent, these novel ubiquitin reagents can be used to capture and subsequently release catalytically active DUBs, whereas existing capturing agents bind irreversibly. These novel reagents allow for the study of these enzymes in their active state under various conditions.

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