Purification of metal-dependent lysine deacetylases with consistently high activity

•A relatively simple, straightforward, and robust protocol for KDAC purification.•Applicable to multiple expression hosts.•High yield of highly active, stable enzymes.•Enhanced preparation consistency for more reliable characterization.

Metal-dependent lysine deacetylases (KDACs) are involved in regulation of numerous biological and disease processes through control of post-translational acetylation. Characterization of KDAC activity and substrate identification is complicated by inconsistent activity of prepared enzyme and a range of multi-step purifications. We describe a simplified protocol based on two-step affinity chromatography. The purification method is appropriate for use regardless of expression host, and we demonstrate purification of several representative members of the KDAC family as well as a selection of mutated variants. The purified proteins are highly active and consistent across preparations.