Issue 5, 2024

Discovery of potent and selective activity-based probes (ABPs) for the deubiquitinating enzyme USP30

Abstract

Ubiquitin-specific protease 30 (USP30) is a deubiquitinating enzyme (DUB) localized at the mitochondrial outer membrane and involved in PINK1/Parkin-mediated mitophagy, pexophagy, BAX/BAK-dependent apoptosis, and IKKβ-USP30-ACLY-regulated lipogenesis/tumorigenesis. A USP30 inhibitor, MTX652, has recently entered clinical trials as a potential treatment for mitochondrial dysfunction. Small molecule activity-based probes (ABPs) for DUBs have recently emerged as powerful tools for in-cell inhibitor screening and DUB activity analysis, and here, we report the first small molecule ABPs (IMP-2587 and IMP-2586) which can profile USP30 activity in cells. Target engagement studies demonstrate that IMP-2587 and IMP-2586 engage active USP30 at nanomolar concentration after only 10 min incubation time in intact cells, dependent on the presence of the USP30 catalytic cysteine. Interestingly, proteomics analyses revealed that DESI1 and DESI2, small ubiquitin-related modifier (SUMO) proteases, can also be engaged by these probes, further suggesting a novel approach to develop DESI ABPs.

Graphical abstract: Discovery of potent and selective activity-based probes (ABPs) for the deubiquitinating enzyme USP30

Supplementary files

Article information

Article type
Paper
Submitted
29 jan 2024
Accepted
07 mar 2024
First published
13 mar 2024
This article is Open Access
Creative Commons BY license

RSC Chem. Biol., 2024,5, 439-446

Discovery of potent and selective activity-based probes (ABPs) for the deubiquitinating enzyme USP30

M. Mondal, F. Cao, D. Conole, H. W. Auner and E. W. Tate, RSC Chem. Biol., 2024, 5, 439 DOI: 10.1039/D4CB00029C

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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