Fluorinated Aromatic Amino Acids Are Sensitive 19F NMR Probes for Bromodomain-Ligand Interactions

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• 作者：Neeraj K. Mishra ; Andrew K. Urick ; Stuart W. J. Ember ; Ernst Sch枚nbrunn ; William C. Pomerantz
• 刊名：ACS Chemical Biology
• 出版年：2014
• 出版时间：December 19, 2014
• 年：2014
• 卷：9
• 期：12
• 页码：2755-2760
• 全文大小：322K
• ISSN：1554-8937

文摘

We describe a ¹⁹F NMR method for detecting bromodomain鈥搇igand interactions using fluorine-labeled aromatic amino acids due to the conservation of aromatic residues in the bromodomain binding site. We test the sensitivity, accuracy, and speed of this method with small molecule ligands (+)-JQ1, BI2536, Dinaciclib, TG101348, and acetaminophen using three bromodomains Brd4, BrdT, and BPTF. Simplified ¹⁹F NMR spectra allowed for simultaneous testing of multiple bromodomains to assess selectivity and identification of a new BPTF ligand. Fluorine labeling only modestly affected the Brd4 structure and function assessed by isothermal titration calorimetry, circular dichroism, and X-ray crystallography. The speed, ease of interpretation, and low concentration of protein needed for binding experiments affords a new method to discover and characterize both native and new ligands.