title: An improved fluorogenic assay for SIRT1, SIRT2, and SIRT3.
abstract: Sirtuins are NAD-dependent lysine deacylases that play critical roles in cellular regulation and are implicated in human diseases. Modulators of sirtuins are needed as tools for investigating their biological functions and possible therapeutic applications. However, the discovery of sirtuin modulators is hampered by the lack of efficient sirtuin assays. Here we report an improved fluorogenic assay for SIRT1, SIRT2, and SIRT3 using a new substrate, a myristoyl peptide with a C-terminal aminocoumarin. The new assay has several advantages, including significantly lower substrate concentration needed, increased signal-to-background ratio, and improved Z'-factor. The novel assay thus will expedite high-throughput screening of SIRT1, SIRT2, and SIRT3 modulators.
authors.affiliation: Department of Chemistry and Chemical Biology, Baker Laboratory, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Ying-Ling
authors.initials: YL
authors.lastname: Chiang
authors.forename: Hening
authors.initials: H
authors.lastname: Lin
mesh_terms: D001681:Biological Assay / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D005453:Fluorescence; D006801:Humans; D020128:Inhibitory Concentration 50; D007700:Kinetics; D015394:Molecular Structure; D009536:Niacinamide / Q000737:chemistry; D037761:Sirtuins / Q000737:chemistry* / Q000145:classification*
chemical_list: D009536:Niacinamide; D037761:Sirtuins