Highly Sensitive Lysine Deacetylase Assay Based on Acetylated Firefly Luciferase
Martin Spinck1, Maria Ecke1, Sonja Sievers1
1Max-Planck-Institute of Molecular Physiology , Otto-Hahn-Strasse 11 , 44227 Dortmund , Germany.
Abstract:
Lysine deacetylases (KDACs) play important roles in many physiological processes and are implicated in many human diseases. Hence, the search for modulators of KDACs is very active, and reliable assays for monitoring their activity are key to success. Here, we describe a new KDAC assay based on Firefly luciferase harboring an acetylation on an essential active site lysine. We show that several KDACs can reverse this modification and hence activate luciferase. This new assay is extremely sensitive, reliable, and fast and can be performed in a continuous format. We used this assay to screen a small library of compounds and identified several novel effectors of SirT2 with low micromolar activity.
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