fedratinib and dinaciclib

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.

Topics: Acetaminophen; Amino Acids, Aromatic; Antigens, Nuclear; Azepines; Binding Sites; Bridged Bicyclo Compounds, Heterocyclic; Cell Cycle Proteins; Crystallography, X-Ray; Cyclic N-Oxides; Escherichia coli; Gene Expression; Halogenation; Humans; Indolizines; Ligands; Magnetic Resonance Spectroscopy; Molecular Docking Simulation; Molecular Probes; Nerve Tissue Proteins; Nuclear Proteins; Oncogene Proteins, Fusion; Protein Binding; Pteridines; Pyridinium Compounds; Pyrrolidines; Recombinant Proteins; Staining and Labeling; Sulfonamides; Transcription Factors; Triazoles