Work

Identification, optimisation, and structural elucidation of chloroacetamide scaffold as covalent inhibitors for ubiquitin C-terminal Hydrolase L3

Year: 2026

Type: article

Abstract: The deubiquitinating enzyme, ubiquitin C-terminal hydrolase L3 (UCHL3), has been implicated as a potential therapeutic target for cancer with a role in regulating the DNA damage response pathways. While the target has been studied using genetic methods there is a lack of reliable chemical probes to selectivity target UCHL3. In this study we report hit identification and optimisation of a new chemical scaffold that irreversibly inhibits UCHL3. The observed structure-activity relationships are corroborated by ligand-bound crystal structures that confirm covalent adduct formation with the catalytic cysteine of the enzyme. Finally, through gel-shift assays using a ubiquitin activity-based probe we demonstrate on-target engagement with UCHL3 in two cell lines. The work as a whole presents a comprehensive evaluation of the new scaffold that can be utilised to probe UCHL3 in different biological contexts.

Source: Figshare

DOI: 10.6084/m9.figshare.34912717

Authors: Nagabhushana C. Beeralingappa, Mengyao Lu, Rishi Patel, Nipuni Pannala, Alisha Dhiman, Brittany N. Heil, Ryan D. Imhoff, Emily G. Smith, Mei B. Bahler, Haley L. Marsden, Brittany L. Allen-Petersen, Michael K. Wendt, Chittaranjan Das, Daniel P. Flaherty

Language: English

Topic: Ubiquitin and proteasome pathways

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

OpenAlex ID: W7225262383

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