Fluorinated Aromatic Amino Acids Are Sensitive 19F NMR Probes for Bromodomain-Ligand Interactions

Autor:	Andrew K. Urick, S.W. Ember, Neeraj K. Mishra, Ernst Schönbrunn, William C. K. Pomerantz
Jazyk:	angličtina
Rok vydání:	2014
Předmět:	Circular dichroism Chemistry Stereochemistry Isothermal titration calorimetry General Medicine Nuclear magnetic resonance spectroscopy Fluorine-19 NMR Biochemistry Small molecule Bromodomain chemistry.chemical_compound Aromatic amino acids Molecular Medicine Letters Binding site
Zdroj:	ACS Chemical Biology
ISSN:	1554-8937 1554-8929
Popis:	We describe a 19F 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 19F 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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ca9c9c27cb9e2758801a9265b3c2a564 http://europepmc.org/articles/PMC4273984 Zobrazit plný text záznamu