Fluorinated Aromatic Amino Acids Are Sensitive 19F NMR Probes for Bromodomain-Ligand Interactions
Year: 2014
Type: article
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.
Source: ACS Chemical Biology
DOI: 10.1021/cb5007344
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $4,500
Funders: University of Minnesota, National Institutes of Health, NIH Office of the Director, National Cancer Institute, National Institute of General Medical Sciences, National Institute of Neurological Disorders and Stroke, Division of Chemistry, Eunice Kennedy Shriver National Institute of Child Health and Human Development
Awards: G1584080787, Drug Discovery & Synthesis of Contraceptive Agents, iTC200 MicroCalorimetry Cell Control Unit, G3058465331, Modeling Pheromone Induced Plasmid and Drug Resistance Transfer, G3293496369, G4678128053, G563102849, G6978267004, G71118468, G7149605548, Moffitt Cancer Center Support Grant
OpenAlex ID: W2082837622