High-Throughput Screening for Chemical Inhibitors of Histone Acetyltransferase

Histone acetyltransferase 1 (HAT1) is an enzyme which acetylates lysine on histone proteins and is intricately involved with regulating gene transcription. Thus, HAT1 is a promising therapeutic target for cancer and a high-throughput method to discover potent inhibitors would be valuable. A fluorescence-based, enzymatic assay can be constructed using a biotinylated HAT1 substrate, recombinant HAT1, and an acetyl co-A substrate mimetic with an alkyne click handle. The HAT1 substrate can be captured by wells coated with streptavidin. Biotin-azide can be conjugated to alkyne-containing lysine through click chemistry. Addition of streptavidin-HRP facilitates the measurement of activity via plate reader.

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