Dataset Information

Discovery of highly Potent and Selective Activity-Based Probes for the Deubiquitylating Enzyme USP30

ABSTRACT: Ubiquitin-specific protease 30 (USP30) is a deubiquitylating enzyme (DUB) localized in the mitochondrial membrane, which is related to PINK1/Parkin-mediated mitophagy, pexophagy, BAX/BAK-dependent apoptosis, and IKKβ-USP30-ACLY-regulated lipogenesis/tumorigenesis. Mission therapeutics pointed their in-house screened MTX652 as USP30 inhibitor for Phase I clinical trial in early 2022. Activity-based probes (ABPs) provide a powerful tool for screen USP30 inhibitors. Here, we report the first small molecule ABPs (ABP 2 and ABP 4) for profiling activity of USP30. Through in-gel fluorescence, target-enrichment and proteomics analysis, we demonstrate that ABP 2 and ABP 4 selectively engage USP30 at nanomolar concentration for only 10 min incubation time in live cells. This cellular USP30-engagement is selectively depending on the catalytic cysteine of USP30. Interestingly, DESI1 and DESI2, the small ubiquitin-related modifier (SUMO) proteases, are also engaged by ABP 2 and ABP 4, providing the novel strategy for these probes as DESIs ABPs. We use proteomics analysis to identify the probes targeted proteins by DIA analysis.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Fangyuan Cao

LAB HEAD: Ed Tate

PROVIDER: PXD044792 | Pride | 2024-05-22

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	Experimental_design_1.txt	Txt
	FC-MM-18-F1_20220704083722.raw	Raw
	FC-MM-18-F2_20220704125234.raw	Raw
	FC-MM-18-F3_20220704170728.raw	Raw
	FC-MM-18-F4_20220704212144.raw	Raw

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Publications

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

Mondal Milon M Cao Fangyuan F Conole Daniel D Auner Holger W HW Tate Edward W EW

RSC chemical biology 20240313 5

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 ...[more]

PMID: 38725909

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