Cell-Based Covalent-Capture Deubiquitinase Assay for Inhibitor Discovery

• DOI: 10.1021/acsptsci.4c00331
• Publication Date: 2024-08-17T01:52:31Z
• Authors: Megan N. Doleschal; Jenna Miller; Sankalp Jain; Alexey V. Zakharov; Ganesha Rai; Anton Simeonov; Bolormaa Baljinnyam; Zhihao Zhuang
• Keywords: Deubiquitinating enzyme

Ubiquitination is a post-translational modification that elicits variety of cellular responses. Deubiquitinases (DUBs) remove ubiquitin moieties from proteins and modulate processes by counteracting the ligase activities. deubiquitination are tightly regulated different mechanisms their dysregulation associated with many diseases. Discovery DUB inhibitors could not only lead to therapeutics but also facilitate understanding ubiquitination/deubiquitination regulatory mechanisms. To enable inhibitor discovery against DUBs, we developed cell-based assay utilizes cell-permeable probe, Biotin-cR