title: Activity-based chemical proteomics accelerates inhibitor development for deubiquitylating enzymes.
abstract: 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: Nuffield Department of Medicine, Henry Wellcome Building for Molecular Physiology, University of Oxford, Oxford OX3 7DQ.
authors: Mikael
authors: M
authors: Altun
authors: Holger B
authors: HB
authors: Kramer
authors: Lianne I
authors: LI
authors: Willems
authors: Jeffrey L
authors: JL
authors: McDermott
authors: Craig A
authors: CA
authors: Leach
authors: Seth J
authors: SJ
authors: Goldenberg
authors: K G Suresh
authors: KG
authors: Kumar
authors: Rebecca
authors: R
authors: Konietzny
authors: Roman
authors: R
authors: Fischer
authors: Edward
authors: E
authors: Kogan
authors: Mukram M
authors: MM
authors: Mackeen
authors: Joanna
authors: J
authors: McGouran
authors: Svetlana V
authors: SV
authors: Khoronenkova
authors: Jason L
authors: JL
authors: Parsons
authors: Grigory L
authors: GL
authors: Dianov
authors: Benjamin
authors: B
authors: Nicholson
authors: Benedikt M
authors: BM
authors: Kessler
mesh_terms: D000631:Aminopyridines / Q000737:chemistry / Q000494:pharmacology*; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D040901:Proteomics; D034622:RNA Interference; D053719:Tandem Mass Spectrometry; D013861:Thiocyanates / Q000737:chemistry / Q000494:pharmacology*; D013876:Thiophenes / Q000737:chemistry / Q000494:pharmacology*; D043222:Ubiquitin Thiolesterase / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D000076084:Ubiquitin-Specific Peptidase 7
chemical_list: C570895:1-(5-((2,4-difluorophenyl)thio)-4-nitrothiophen-2-yl)ethanone; C570894:2,6-diaminopyridine-3,5-bis(thiocyanate); D000631:Aminopyridines; D000906:Antibodies; D004791:Enzyme Inhibitors; D013861:Thiocyanates; D013876:Thiophenes; C464569:USP7 protein, human; D043222:Ubiquitin Thiolesterase; D000076084:Ubiquitin-Specific Peptidase 7