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.