Fluorinated aromatic amino acids are sensitive 19 f NMR probes for bromodomain-ligand interactions

Neeraj K. Mishra, Andrew K. Urick, Stuart W.J. Ember, Ernst Schonbrunn, William C. Pomerantz

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

We describe a 19 F NMR method for detecting bromodomain-ligand 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 19 F 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.

Original languageEnglish (US)
Pages (from-to)2755-2760
Number of pages6
JournalACS Chemical Biology
Volume9
Issue number12
DOIs
StatePublished - Dec 19 2014

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Aromatic Amino Acids
Nuclear magnetic resonance
Ligands
Fluorine
Calorimetry
X ray crystallography
X Ray Crystallography
Dichroism
Acetaminophen
Circular Dichroism
Titration
Protein Binding
Labeling
Conservation
Binding Sites
Molecules
Testing
Proteins
Experiments

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Fluorinated aromatic amino acids are sensitive 19 f NMR probes for bromodomain-ligand interactions . / Mishra, Neeraj K.; Urick, Andrew K.; Ember, Stuart W.J.; Schonbrunn, Ernst; Pomerantz, William C.

In: ACS Chemical Biology, Vol. 9, No. 12, 19.12.2014, p. 2755-2760.

Research output: Contribution to journalArticle

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