title: Quantitative Chemical Proteomic Profiling of Ubiquitin Specific Proteases in Intact Cancer Cells.
abstract: Deubiquitinating enzymes play an important role in a plethora of therapeutically relevant processes and are emerging as pioneering drug targets. Herein, we present a novel probe, Ubiquitin Specific Protease (USP) inhibitor, alongside an alkyne-tagged activity-based probe analogue. Activity-based proteome profiling identified 12 USPs, including USP4, USP16, and USP33, as inhibitor targets using submicromolar probe concentrations. This represents the first intact cell activity-based profiling of deubiquitinating enzymes. Further analysis demonstrated functional inhibition of USP33 and identified a synergistic relationship in combination with ATR inhibition, consistent with USP4 inhibition.
authors: Department of Chemistry, Imperial College London , London, SW7 2AZ, United Kingdom.
authors: Jennifer A
authors: JA
authors: Ward
authors: MISSION Therapeutics, Ltd. , Moneta, Babraham Research Campus, Cambridge, CB22 3AT, United Kingdom.
authors: Lauren
authors: L
authors: McLellan
authors: MISSION Therapeutics, Ltd. , Moneta, Babraham Research Campus, Cambridge, CB22 3AT, United Kingdom.
authors: Martin
authors: M
authors: Stockley
authors: Sandexis Medicinal Chemistry, Ltd. , Innovation House, Discovery Park, Ramsgate Road, Sandwich, Kent, CT13 9ND, United Kingdom.
authors: Karl R
authors: KR
authors: Gibson
authors: Sandexis Medicinal Chemistry, Ltd. , Innovation House, Discovery Park, Ramsgate Road, Sandwich, Kent, CT13 9ND, United Kingdom.
authors: Gavin A
authors: GA
authors: Whitlock
authors: MISSION Therapeutics, Ltd. , Moneta, Babraham Research Campus, Cambridge, CB22 3AT, United Kingdom.
authors: Charlotte
authors: C
authors: Knights
authors: MISSION Therapeutics, Ltd. , Moneta, Babraham Research Campus, Cambridge, CB22 3AT, United Kingdom.
authors: Jeanine A
authors: JA
authors: Harrigan
authors: MISSION Therapeutics, Ltd. , Moneta, Babraham Research Campus, Cambridge, CB22 3AT, United Kingdom.
authors: Xavier
authors: X
authors: Jacq
authors: Department of Chemistry, Imperial College London , London, SW7 2AZ, United Kingdom.
authors: Edward W
authors: EW
authors: Tate
mesh_terms: D000480:Alkynes / Q000737:chemistry; D045744:Cell Line, Tumor; D006801:Humans; D015336:Molecular Probe Techniques; D015335:Molecular Probes / Q000737:chemistry*; D009369:Neoplasms / Q000201:enzymology*; D040901:Proteomics / Q000379:methods*; D011758:Pyrroles / Q000737:chemistry*; D054852:Small Molecule Libraries / Q000737:chemistry*; D064570:Ubiquitin-Specific Proteases / Q000032:analysis* / Q000037:antagonists & inhibitors
chemical_list: D000480:Alkynes; D015335:Molecular Probes; D011758:Pyrroles; D054852:Small Molecule Libraries; C051267:2-acetylpyrrole; D064570:Ubiquitin-Specific Proteases