TY - JOUR
T1 - Fluorinated aromatic amino acids are sensitive 19 f NMR probes for bromodomain-ligand interactions
AU - Mishra, Neeraj K.
AU - Urick, Andrew K.
AU - Ember, Stuart W.J.
AU - Schonbrunn, Ernst
AU - Pomerantz, William C.
N1 - Publisher Copyright:
© 2014 American Chemical Society.
PY - 2014/12/19
Y1 - 2014/12/19
N2 - 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.
AB - 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.
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U2 - 10.1021/cb5007344
DO - 10.1021/cb5007344
M3 - Article
C2 - 25290579
AN - SCOPUS:84919675043
SN - 1554-8929
VL - 9
SP - 2755
EP - 2760
JO - ACS Chemical Biology
JF - ACS Chemical Biology
IS - 12
ER -