title: Fluorinated aromatic amino acids are sensitive 19F NMR probes for bromodomain-ligand interactions.
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.
authors.affiliation: Department of Chemistry, University of Minnesota , 207 Pleasant St. SE, Minneapolis, Minnesota 55455, United States.
authors.forename: Neeraj K
authors.initials: NK
authors.lastname: Mishra
authors.forename: Andrew K
authors.initials: AK
authors.lastname: Urick
authors.forename: Stuart W J
authors.initials: SW
authors.lastname: Ember
authors.forename: Ernst
authors.initials: E
authors.lastname: Schönbrunn
authors.forename: William C
authors.initials: WC
authors.lastname: Pomerantz
mesh_terms: D000082:Acetaminophen / Q000737:chemistry; D024322:Amino Acids, Aromatic / Q000737:chemistry*; D034961:Antigens, Nuclear / Q000737:chemistry / Q000235:genetics; D001381:Azepines / Q000737:chemistry; D001665:Binding Sites; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000737:chemistry; D018797:Cell Cycle Proteins; D018360:Crystallography, X-Ray; D003497:Cyclic N-Oxides; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D054879:Halogenation; D006801:Humans; D007212:Indolizines; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D062105:Molecular Docking Simulation; D015335:Molecular Probes / Q000737:chemistry*; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics; D015514:Oncogene Proteins, Fusion / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding; D011621:Pteridines / Q000737:chemistry; D011726:Pyridinium Compounds / Q000737:chemistry; D011759:Pyrrolidines / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013194:Staining and Labeling / Q000379:methods; D013449:Sulfonamides / Q000737:chemistry; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics; D014230:Triazoles / Q000737:chemistry; D000092468:Bromodomain Containing Proteins; D000096926:Benzenesulfonamides
chemical_list: D000082:Acetaminophen; D024322:Amino Acids, Aromatic; D034961:Antigens, Nuclear; D001381:Azepines; D019086:Bridged Bicyclo Compounds, Heterocyclic; D018797:Cell Cycle Proteins; D003497:Cyclic N-Oxides; D007212:Indolizines; D008024:Ligands; D015335:Molecular Probes; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D015514:Oncogene Proteins, Fusion; D011621:Pteridines; D011726:Pyridinium Compounds; D011759:Pyrrolidines; D011994:Recombinant Proteins; D013449:Sulfonamides; D014157:Transcription Factors; D014230:Triazoles; C561695:(+)-JQ1 compound; C518477:BI 2536; C441289:BRD4 protein, human; D000092468:Bromodomain Containing Proteins; C109570:BRDT protein, human; C406238:fetal Alzheimer antigen; C553669:dinaciclib; C528327:fedratinib; D000096926:Benzenesulfonamides