Dual Labeling of the CBP/p300 KIX Domain for ¹⁹ F NMR Leads to Identification of a New Small-Molecule Binding Site.

ABSTRACT: Protein-Observed Fluorine NMR (PrOF NMR) spectroscopy is an emerging technique for screening and characterizing small-molecule-protein interactions. The choice of which amino acid to label for PrOF NMR can be critical for analysis. Here we report the first use of a protein containing two different fluoroaromatic amino acids for NMR studies. Using the KIX domain of the CBP/p300 as a model system, we examine ligand binding of several small-molecule compounds elaborated from our previous fragment screen and identify a new ligand binding site distinct from those used by native transcription factors. This site was further supported by computational modeling (FTMap and Schrödinger) and ¹ H,¹⁵ N HSQC/HMQC NMR spectroscopy. Metabolic labeling with multiple fluorinated amino a

SUBMITTER: Gee CT 

PROVIDER: S-EPMC6251716 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature