Protein-Observed Fluorine NMR Is a Complementary Ligand Discovery Method to ¹H CPMG Ligand-Observed NMR.

ABSTRACT: To evaluate its potential as a ligand discovery tool, we compare a newly developed 1D protein-observed fluorine NMR (PrOF NMR) screening method with the well-characterized ligand-observed ¹H CPMG NMR screen. We selected the first bromodomain of Brd4 as a model system to benchmark PrOF NMR because of the high ligandability of Brd4 and the need for small molecule inhibitors of related epigenetic regulatory proteins. We compare the two methods' hit sensitivity, triaging ability, experiment speed, material consumption, and the potential for false positives and negatives. To this end, we screened 930 fragment molecules against Brd4 in mixtures of five and followed up these studies with mixture deconvolution and affinity characterization of the top hits. In selected examples, we also

SUBMITTER: Urick AK 

PROVIDER: S-EPMC8325173 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature