The Sirtuin family of histone/protein deacetylating enzymes has gained considerable interest both for their recognized importance in processes such as gene silencing and expression and in cellular metabolism. This has led to identification of these enzymes as important actors in diverse pathologies including diabetes, Alzheimer's disease and oncology. We have developed a convenient and sensitive screening methodology to detect and measure both inhibitors and activators of sirtuin activity. Here we describe a one-step, homogeneous luminescent assay for the detection of activity for multiple sirtuin isotypes. The reagent formulation contains a pro-luminescent sirtuin peptide substrate, NAD+, ATP, thermostable luciferase, and a developer enzyme that cleaves deacetylated lysine residues. The addition of an active sirtuin enzyme source leads to a robust luminescent signal within minutes with “glow-type” characteristics lasting for several hours. Data will be presented showing characterization of the assay using a diverse assortment of established sirtuin inhibitors, followed by suitability for screening in 384-well formats and the incorporation of a control substrate to identify false positives.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4050. doi:10.1158/1538-7445.AM2011-4050