Design and Application of Cofactor Analogues as Chemical Tools to Probe Lysine Acetyltransferase Ligands Targeting the Catalytic Domain

Here we present the synthesis of two functional cofactor-based chemical probes and their usage as mechanistic tools in a broadly applica-ble assay platform for histone acetyltransferases (HATs). In this platform, a fluorescence polarization (FP) based binding assay was combined with biolayer interferometry (BLI) competition analysis and a FP competition activity immunoassay to enable easy, reliable, and pro-found evaluation of ligands that target the KAT cofactor binding site.