Integration of Bioorthogonal Probes and Q-FRET for the Detection of Histone Acetyltransferase Activity

摘要

Histone acetyltransferases (HATs) are key players in epigenetic regulation of gene function. Recent discovery of diverse HAT substrates implicates a broad spectrum of cellular functions of HATs. Many pathological processes are also intimately associated with dysregulation of HAT levels and activities. However, detection of enzymatic activity of HATs has been challenging and significantly impeded drug discovery. To advance the field, we developed a convenient one-pot mix-and-read strategy that is capable to directly detect the acylated histone product via fluorescent readout. The strategy integrated three technological platforms, bioorthogonal HAT substrate labeling, alkyne-azide click chemistry, and quenching-FRET, into one system for effective probing of HAT enzyme activity.