title: Discovery of SIRT3 Inhibitors Using SAMDI Mass Spectrometry.
abstract: Lysine acetylation plays a critical role in cellular regulation and is implicated in human disease. Sirtuin deacetylases remove acetyl groups from modified lysine residues, and sirtuin 3 (SIRT3) has been identified as a target for cancer therapeutics. Robust and high-throughput screening methods for these targets will be important to the development of therapeutics. This article describes the use of self-assembled monolayer desorption/ionization mass spectrometry, or SAMDI-MS-a label-free drug discovery tool--to characterize SIRT3 activity and discover inhibitors. SAMDI-MS was used to analyze a peptide array having 361 distinct acetylated peptides to identify an active SIRT3 substrate (GYK(Ac)RGC). This peptide was used in a screen of 100,000 small molecules to identify inhibitors of SIRT3. A total of 306 SIRT3 inhibitors were identified, with one compound, SDX-437, having an IC(50) of 700 nM with >100-fold selectivity for SIRT3 over SIRT1.
authors.affiliation: SAMDI Tech, Inc., Chicago, IL, USA.
authors.forename: Kaushal
authors.initials: K
authors.lastname: Patel
authors.affiliation: SAMDI Tech, Inc., Chicago, IL, USA.
authors.forename: John
authors.initials: J
authors.lastname: Sherrill
authors.affiliation: Department of Biomedical Engineering, Department of Chemistry, and Department of Cell & Molecular Biology, Northwestern University, Evanston, IL, USA.
authors.forename: Milan
authors.initials: M
authors.lastname: Mrksich
authors.affiliation: SAMDI Tech, Inc., Chicago, IL, USA mscholle@samditech.com.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Scholle
mesh_terms: D000107:Acetylation; D055808:Drug Discovery / Q000379:methods; D004789:Enzyme Activation / Q000187:drug effects; D057166:High-Throughput Screening Assays; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D056566:Sirtuin 3 / Q000037:antagonists & inhibitors*
chemical_list: D056572:Histone Deacetylase Inhibitors; D056566:Sirtuin 3
keywords: deacetylase; label-free; peptide array; self-assembled monolayers