Dual Screening of BPTF and Brd4 Using Protein-Observed Fluorine NMR Uncovers New Bromodomain Probe Molecules

Urick, A. K., Hawk, L. M., Cassel, M. K., Mishra, N. K., Liu, S., Adhikari, N., Zhang, W., Dos Santos, C. O., Hall, J. L., Pomerantz, W. C. (October 2015) Dual Screening of BPTF and Brd4 Using Protein-Observed Fluorine NMR Uncovers New Bromodomain Probe Molecules. ACS Chem Biol, 10 (10). pp. 2246-56. ISSN 1554-8937 (Electronic)1554-8929 (Linking)

URL: http://www.ncbi.nlm.nih.gov/pubmed/26158404
DOI: 10.1021/acschembio.5b00483

Abstract

Bromodomain-containing protein dysregulation is linked to cancer, diabetes, and inflammation. Selective inhibition of bromodomain function is a newly proposed therapeutic strategy. We describe a (19)F NMR dual screening method for small molecule discovery using fluorinated tryptophan resonances on two bromodomain-containing proteins. The chemical shift dispersion of (19)F resonances within fluorine-labeled proteins enables the simultaneous analysis of two fluorinated bromodomains by NMR. A library of 229 small molecules was screened against the first bromodomain of Brd4 and the BPTF bromodomain. We report the first small molecule selective for BPTF over Brd4, termed AU1. The Kd = 2.8 muM for AU1, which is active in a cell-based reporter assay. No binding is detected with Brd4. Three new Brd4 inhibitors with submicromolar affinity were also discovered. Brd4 hits were validated in a thermal stability assay and potency determined via fluorescence anisotropy. The speed, ease of interpretation, and low protein concentration needed for protein-observed (19)F NMR experiments in a multiprotein format offers a new method to discover and characterize selective ligands for bromodomain-containing proteins.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > BET bromodomain coactivator protein
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > BET bromodomain coactivator protein > Brd4
diseases & disorders > cancer > drugs and therapies
Investigative techniques and equipment > spectroscopy > magnetic resonance spectroscopy
CSHL Authors:
Communities: CSHL labs > Dos Santos lab
CSHL Cancer Center Program > Gene Regulation and Cell Proliferation
Depositing User: Matt Covey
Date: 16 October 2015
Date Deposited: 09 Nov 2015 17:44
Last Modified: 06 Mar 2018 16:56
PMCID: PMC4858447
Related URLs:
URI: http://repository.cshl.edu/id/eprint/31965

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