A Flexible and Robust Solution for Automated 384-well HDAC Profiling

Authors: Brad Larson, Peter Banks, BioTek Instruments, Inc., Winooski, Vermont; Diana Hulboy, Kara Cannon , Wayne Patton, Enzo Life Sciences, Farmingdale, New York

Histone deacetylases (HDACs) are a class of enzymes that remove acetyl groups from ε-N-acetyl lysine amino acids on histones and other proteins. The removal of acetyl groups serves to increase the positive charge of histone tails, encouraging binding between histones and the DNA backbone, and preventing transcription. Because of this role, HDACs have been implicated in a variety of human diseases, including cancer; making them an increasingly popular target for drug discovery research. Here we demonstrate an application to monitor the deacetylation activity of multiple Class I and II HDAC enzymes in an automated fashion. Validation and pharmacology results demonstrate how the fluorescent assay technology and instrumentation can be used together to provide a relevant system to assess enzyme kinetics and inhibition.