title: Identification of the Binding Site of Chroman-4-one-Based Sirtuin 2-Selective Inhibitors using Photoaffinity Labeling in Combination with Tandem Mass Spectrometry.
abstract: Photoaffinity labeling (PAL) was used to identify the binding site of chroman-4-one-based SIRT2-selective inhibitors. The photoactive diazirine 4, a potent SIRT2 inhibitor, was subjected to detailed photochemical characterization. In PAL experiments with SIRT2, a tryptic peptide originating from the covalent attachment of photoactivated 4 was identified. The peptide covers both the active site of SIRT2 and the proposed binding site of chroman-4-one-based inhibitors. A high-power LED was used as source for the monochromatic UV light enabling rapid photoactivation.
authors.affiliation: Department of Chemistry and Molecular Biology, Medicinal Chemistry, University of Gothenburg , Kemivagen 10, SE-41296 Göteborg, Sweden.
authors.forename: Tina
authors.initials: T
authors.lastname: Seifert
authors.affiliation: Department of Chemistry and Molecular Biology, Medicinal Chemistry, University of Gothenburg , Kemivagen 10, SE-41296 Göteborg, Sweden.
authors.forename: Marcus
authors.initials: M
authors.lastname: Malo
authors.affiliation: The Proteomics Core Facility, Sahlgrenska Academy, University of Gothenburg , SE-41390 Göteborg, Sweden.
authors.forename: Johan
authors.initials: J
authors.lastname: Lengqvist
authors.affiliation: The Proteomics Core Facility, Sahlgrenska Academy, University of Gothenburg , SE-41390 Göteborg, Sweden.
authors.forename: Carina
authors.initials: C
authors.lastname: Sihlbom
authors.affiliation: School of Pharmacy, University of Eastern Finland , P.O. Box 1627, 70211 Kuopio, Finland.
authors.forename: Elina M
authors.initials: EM
authors.lastname: Jarho
authors.affiliation: Department of Chemistry and Molecular Biology, Medicinal Chemistry, University of Gothenburg , Kemivagen 10, SE-41296 Göteborg, Sweden.
authors.forename: Kristina
authors.initials: K
authors.lastname: Luthman
mesh_terms: D001665:Binding Sites / Q000187:drug effects; D002867:Chromones / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D008958:Models, Molecular; D015394:Molecular Structure; D019920:Photoaffinity Labels / Q000737:chemistry*; D056565:Sirtuin 2 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013329:Structure-Activity Relationship; D053719:Tandem Mass Spectrometry
chemical_list: D002867:Chromones; D004791:Enzyme Inhibitors; D019920:Photoaffinity Labels; C068514:4-chromanone; C474199:SIRT2 protein, human; D056565:Sirtuin 2