Poster: Homogenous detection of O-Acetyl-ADP-ribose as a generic HTS assay method for SIRTs

Sirtuins (SIRTs) are deacetylases that catalyze removal of acetyl groups from histones and other proteins; they are important components of the epigenetic regulatory landscape and have been targeted for diverse indications, including cancer.  SIRTs deacetylate in an NAD-dependent reaction that yields O-acetyl-ADP-ribose (OAADPr) and nicotinamide as products along with the deacetylated protein.  Screening and hit-to-lead efforts targeting SIRTs has been hampered by a lack of robust HTS assays methods for detection of SIRT enzymatic activity, a situation that is exacerbated by the diversity of deacetylated protein products and the inherent instability of the OAADPr product.  To address this unmet need, we developed a generic activity assay for SIRTs based on enzymatic conversion of OAADPr to AMP in real time, which is then detected using the Transcreener AMP/GMP assay with a fluorescence polarization (FP) or time resolved Förster resonance energy transfer (TR-FRET) readout.