Format

Send to

Choose Destination
ACS Chem Biol. 2015 Oct 16;10(10):2246-56. doi: 10.1021/acschembio.5b00483. Epub 2015 Jul 28.

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

Author information

1
Department of Chemistry, University of Minnesota , 207 Pleasant St. SE, Minneapolis, Minnesota 55455, United States.
2
Department of Chemistry, University of Massachusetts Boston , 100 Morrissey Boulevard, Boston, Massachusetts 02125, United States.
3
Lillehei Heart Institute , Department of Medicine, 2231 6th Street SE, Minneapolis, Minnesota 55455, United States.
4
Cold Spring Harbor Laboratory , 1 Bungtown Road, Cold Spring Harbor, New York 11724, United States.

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 μM 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.

PMID:
26158404
PMCID:
PMC4858447
DOI:
10.1021/acschembio.5b00483
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for American Chemical Society Icon for PubMed Central
Loading ...
Support Center