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ID: ALA4808148
Title: HDAC6 screening dataset using tau-based substrate in an enzymatic assay yields selective inhibitors and activators
Authors: Bernhard Ellinger, Justus Dick, Vanessa Lage-Rupprecht, Bruce Schultz, Andrea Zaliani, Marcin Namysl, Stephan Gebel, Ole Pless, Jeanette Reinshagen, Christian Ebeling, Alexander Esser, Marc Jacobs, Carsten Claussen, and Martin Hofmann-Apitius
Abstract: Histone deacetylase 6 (HDAC6) and HDAC10 are unique among the other HDACs as they consist of two catalytically active domains instead of one. Only in case of HDAC6 both domains are active resulting in a number of unique deacetylase reactions. HDAC6 can regulate the microtubule network and plays a role in the degradation of misfolded and aggregated proteins, making it a relevant target for respective disease indications. We therefore developed a substrate (Boc-Ile-Asp-(Dimethyl)Lys-(Ac)Lys-aminoluciferin) based on a critical acetylation site of misfolded human MAPT/Tau, a hallmark of Alzheimer disease. This substrate was used to screen a 5632 compound containing repurposing library at 10 �M in a coupled luminescence based assay. The assay was miniaturised to 10 �L per enzymatic reaction. For comparison, a generic HDAC substrate (BOC-Gly-(Ac)Lys-aminoluciferin) was used to screen the same library. Both substrates rely on a cascade of enzymatic reactions. First, HDAC6 deacetylates the substrate followed by cleavage of aminoluciferin from the peptide by porcine trypsin and conversion of the aminoluciferin by Firefly luciferase. Compounds with an activity of at least 75% inhibition for the MAPT/Tau-based substrate were confirmed in triplicates at the screening concentration of 10 �M. Confirmed hits, where analysed in eight or 15 point dose response curves, depending on their activity. The data presented here include both primary data sets with 5632 compounds each as well as 249 values from hit confirmation screening against the MAPT/Tau-based substrate and 151 IC50 values from the confirmed hits.