Activity-based chemical proteomics accelerates inhibitor development for deubiquitylating enzymes.

Journal article

Converting lead compounds into drug candidates is a crucial step in drug development, requiring early assessment of potency, selectivity, and off-target effects. We have utilized activity-based chemical proteomics to determine the potency and selectivity of deubiquitylating enzyme (DUB) inhibitors in cell culture models. Importantly, we characterized the small molecule PR-619 as a broad-range DUB inhibitor, and P22077 as a USP7 inhibitor with potential for further development as a chemotherapeutic agent in cancer therapy. A striking accumulation of polyubiquitylated proteins was observed after both selective and general inhibition of cellular DUB activity without direct impairment of proteasomal proteolysis. The repertoire of ubiquitylated substrates was analyzed by tandem mass spectrometry, identifying distinct subsets for general or specific inhibition of DUBs. This enabled identification of previously unknown functional links between USP7 and enzymes involved in DNA repair.

Authors: Altun, M; Kramer, H; Willems, L; McDermott, J; Leach, C

• Publication status: Published
• Journal: Chemistry and biology
• Volume: 18
• Issue: 11
• Pages: 1401-1412
• Publication date: 2011-11-01
• DOI: 10.1016/j.chembiol.2011.08.018
• EISSN: 1879-1301
• ISSN: 1074-5521
• Language: English
• Keywords: Cell Line; Chromatography, High Pressure Liquid; Proteomics; RNA Interference; Antibodies; Tandem Mass Spectrometry; Thiocyanates; Thiophenes; Enzyme Activation; Enzyme Inhibitors; Ubiquitin Thiolesterase; Aminopyridines; Humans