title: Fluorescence-based active site probes for profiling deubiquitinating enzymes.
abstract: Novel ubiquitin-based active site probes including a fluorescent tag have been developed and evaluated. A new, functionalizable electrophilic trap is utilized allowing for late stage diversification of the probe. Attachment of fluorescent dyes allowed direct detection of endogenous deubiquitinating enzyme (DUB) activities in cell extracts by in-gel fluorescence imaging.
authors.affiliation: Nuffield Department of Medicine, University of Oxford, Henry Welcome Building for Molecular Physiology, Roosevelt Drive, Oxford, OX3 7BN, UK.
authors.forename: Joanna F
authors.initials: JF
authors.lastname: McGouran
authors.forename: Holger B
authors.initials: HB
authors.lastname: Kramer
authors.forename: Mukram M
authors.initials: MM
authors.lastname: Mackeen
authors.forename: Katalin
authors.initials: K
authors.lastname: di Gleria
authors.forename: Mikael
authors.initials: M
authors.lastname: Altun
authors.forename: Benedikt M
authors.initials: BM
authors.lastname: Kessler
mesh_terms: D020134:Catalytic Domain; D057930:Click Chemistry; D010450:Endopeptidases / Q000737:chemistry* / Q000378:metabolism*; D057075:Enzyme Assays / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D008958:Models, Molecular; D025801:Ubiquitin / Q000737:chemistry / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D025801:Ubiquitin; D010450:Endopeptidases