Cookies on this website
We use cookies to ensure that we give you the best experience on our website. If you click 'Continue' we will assume that you are happy to receive all cookies and you will not see this message again. Click 'Find out more' for information on how to change your cookie settings.

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.

Original publication

DOI

10.1016/j.chembiol.2011.08.018

Type

Journal article

Journal

Chem Biol

Publication Date

23/11/2011

Volume

18

Pages

1401 - 1412

Keywords

Aminopyridines, Antibodies, Cell Line, Chromatography, High Pressure Liquid, Enzyme Activation, Enzyme Inhibitors, Humans, Proteomics, RNA Interference, Tandem Mass Spectrometry, Thiocyanates, Thiophenes, Ubiquitin Thiolesterase