title: Structure-Based Development of an Affinity Probe for Sirtuin 2.
abstract: Sirtuins are NAD(+)-dependent protein deacylases that cleave off acetyl groups, as well as other acyl groups, from the ɛ-amino group of lysines in histones and other substrate proteins. Dysregulation of human Sirt2 activity has been associated with the pathogenesis of cancer, inflammation, and neurodegeneration, thus making Sirt2 a promising target for pharmaceutical intervention. Here, based on a crystal structure of Sirt2 in complex with an optimized sirtuin rearranging ligand (SirReal) that shows improved potency, water solubility, and cellular efficacy, we present the development of the first Sirt2-selective affinity probe. A slow dissociation of the probe/enzyme complex offers new applications for SirReals, such as biophysical characterization, fragment-based screening, and affinity pull-down assays. This possibility makes the SirReal probe an important tool for studying sirtuin biology.
authors.affiliation: Institut für Pharmazeutische Wissenschaften, Albert-Ludwigs-Universität Freiburg, Albertstrasse 25, 79104, Freiburg im Breisgau, Germany.
authors.forename: Matthias
authors.identifier: http://orcid.org/0000-0001-7365-3617
authors.initials: M
authors.lastname: Schiedel
authors.affiliation: Institut für Pharmazeutische Wissenschaften, Albert-Ludwigs-Universität Freiburg, Albertstrasse 25, 79104, Freiburg im Breisgau, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Rumpf
authors.affiliation: Institute für Pharmazie, Martin-Luther-Universität Halle-Wittenberg, Wolfgang-Langenbeck-Strasse 4, 06120, Halle, Saale, Germany.
authors.forename: Berin
authors.initials: B
authors.lastname: Karaman
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Magyar Tudósok körútja 2, H-1117, Budapest, Hungary.
authors.forename: Attila
authors.initials: A
authors.lastname: Lehotzky
authors.affiliation: Institute für Biochemie und BIOSS Centre for Biological, Signaling Studies, Albert-Ludwigs-Universität Freiburg, Albertstrasse 21, 79104, Freiburg im Breisgau, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Gerhardt
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Magyar Tudósok körútja 2, H-1117, Budapest, Hungary.
authors.forename: Judit
authors.initials: J
authors.lastname: Ovádi
authors.affiliation: Institute für Pharmazie, Martin-Luther-Universität Halle-Wittenberg, Wolfgang-Langenbeck-Strasse 4, 06120, Halle, Saale, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Sippl
authors.affiliation: Institute für Biochemie und BIOSS Centre for Biological, Signaling Studies, Albert-Ludwigs-Universität Freiburg, Albertstrasse 21, 79104, Freiburg im Breisgau, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Einsle
authors.affiliation: Institut für Pharmazeutische Wissenschaften, Albert-Ludwigs-Universität Freiburg, Albertstrasse 25, 79104, Freiburg im Breisgau, Germany. manfred.jung@pharmazie.uni-freiburg.de.
authors.forename: Manfred
authors.identifier: http://orcid.org/0000-0002-6361-7716
authors.initials: M
authors.lastname: Jung
mesh_terms: D018360:Crystallography, X-Ray; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D015335:Molecular Probes / Q000032:analysis* / Q000138:chemical synthesis / Q000737:chemistry*; D015394:Molecular Structure; D056565:Sirtuin 2 / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D012995:Solubility; D013329:Structure-Activity Relationship
chemical_list: D008024:Ligands; D015335:Molecular Probes; C474199:SIRT2 protein, human; D056565:Sirtuin 2
keywords: deacylases; drug design; protein modifications; proteomics; sirtuins