title: A Continuous, Fluorogenic Sirtuin 2 Deacylase Assay: Substrate Screening and Inhibitor Evaluation.
abstract: Sirtuins are important regulators of lysine acylation, which is implicated in cellular metabolism and transcriptional control. This makes the sirtuin class of enzymes interesting targets for development of small molecule probes with pharmaceutical potential. To achieve detailed profiling and kinetic insight regarding sirtuin inhibitors, it is important to have access to efficient assays. In this work, we report readily synthesized fluorogenic substrates enabling enzyme-economical evaluation of SIRT2 inhibitors in a continuous assay format as well as evaluation of the properties of SIRT2 as a long chain deacylase enzyme. Novel enzymatic activities of SIRT2 were thus established in vitro, which warrant further investigation, and two known inhibitors, suramin and SirReal2, were profiled against substrates containing ε-N-acyllysine modifications of varying length.
authors.affiliation: Center for Biopharmaceuticals and Department of Drug Design & Pharmacology, University of Copenhagen , Universitetsparken 2, DK-2100, Copenhagen, Denmark.
authors.forename: Iacopo
authors.initials: I
authors.lastname: Galleano
authors.affiliation: Institute of Pharmaceutical Sciences, Albert-Ludwigs-Universität Freiburg , Albertstraße 25, 79104 Freiburg im Breisgau, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Schiedel
authors.affiliation: Institute of Pharmaceutical Sciences, Albert-Ludwigs-Universität Freiburg , Albertstraße 25, 79104 Freiburg im Breisgau, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Jung
authors.affiliation: Center for Biopharmaceuticals and Department of Drug Design & Pharmacology, University of Copenhagen , Universitetsparken 2, DK-2100, Copenhagen, Denmark.
authors.forename: Andreas S
authors.initials: AS
authors.lastname: Madsen
authors.affiliation: Center for Biopharmaceuticals and Department of Drug Design & Pharmacology, University of Copenhagen , Universitetsparken 2, DK-2100, Copenhagen, Denmark.
authors.forename: Christian A
authors.initials: CA
authors.lastname: Olsen
mesh_terms: D000081:Acetamides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives / Q000378:metabolism*; D008958:Models, Molecular; D015394:Molecular Structure; D056565:Sirtuin 2 / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013379:Substrate Specificity / Q000187:drug effects; D013498:Suramin / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D013844:Thiazoles / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D000081:Acetamides; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; C000609012:SirReal2; D013844:Thiazoles; D013498:Suramin; C474199:SIRT2 protein, human; D056565:Sirtuin 2; D008239:Lysine