title: Integration of Bioorthogonal Probes and Q-FRET for the Detection of Histone Acetyltransferase Activity.
abstract: Histone acetyltransferases (HATs) are key players in the epigenetic regulation of gene function. The recent discovery of diverse HAT substrates implies a broad spectrum of cellular functions of HATs. Many pathological processes are also intimately associated with the dysregulation of HAT levels and activities. However, detecting the enzymatic activity of HATs has been challenging, and this has significantly impeded drug discovery. To advance the field, we developed a convenient one-pot, mix-and-read strategy that is capable of directly detecting the acylated histone product through a fluorescent readout. The strategy integrates three technological platforms-bioorthogonal HAT substrate labeling, alkyne-azide click chemistry, and quenching FRET-into one system for effective probing of HAT enzyme activity.
authors.affiliation: Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, The University of Georgia, 250 W Green Street, Athens, GA, 30602, USA.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Han
authors.affiliation: Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, The University of Georgia, 250 W Green Street, Athens, GA, 30602, USA.|Department of Medicinal Chemistry, School of Pharmacy, Qingdao University, Qingdao, Shandong, 266000, China.
authors.forename: Yepeng
authors.initials: Y
authors.lastname: Luan
authors.affiliation: Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, The University of Georgia, 250 W Green Street, Athens, GA, 30602, USA. yzheng@uga.edu.
authors.forename: Yujun George
authors.initials: YG
authors.lastname: Zheng
mesh_terms: D000480:Alkynes / Q000737:chemistry; D001386:Azides / Q000737:chemistry; D002384:Catalysis; D057930:Click Chemistry; D003300:Copper / Q000737:chemistry; D061565:Cycloaddition Reaction; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D051548:Histone Acetyltransferases / Q000032:analysis*; D007700:Kinetics
chemical_list: D000480:Alkynes; D001386:Azides; D005456:Fluorescent Dyes; D003300:Copper; D051548:Histone Acetyltransferases
keywords: FRET; HAT activity; bioorthogonal chemical probes; click reaction; protein acetylation