Open AccessQuantifying the Selectivity of Protein–Protein and Small Molecule Interactions with Fluorinated Tandem Bromodomain Reader Proteins
Author(s) -
Prakriti Kalra,
Logan McGraw,
Jennifer R Kimbrough,
Anil K. Pandey,
Jonathan Solberg,
Huarui Cui,
Anand Divakaran,
Kristen John,
Jon E. Hawkinson,
William C. K. Pomerantz
Publication year - 2020
Publication title -
acs chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.899
H-Index - 111
eISSN - 1554-8937
pISSN - 1554-8929
DOI - 10.1021/acschembio.0c00720
Subject(s) - bromodomain , tandem , chemistry , computational biology , protein–protein interaction , small molecule , selectivity , biochemistry , biophysics , biology , epigenetics , gene , materials science , composite material , catalysis
Multidomain bromodomain-containing proteins regulate gene expression via chromatin binding, interactions with the transcriptional machinery, and by recruiting enzymatic activity. Selective inhibition of members of the bromodomain and extra-terminal (BET) family is important to understand their role in disease and gene regulation, although due to the similar binding sites of BET bromodomains, selective inhibitor discovery has been challenging. To support the bromodomain inhibitor discovery process, here we report the first application of protein-observed fluorine (PrOF) NMR to the tandem bromodomains of BRD4 and BRDT to quantify the selectivity of their interactions with acetylated histones as well as small molecules. We further determine the selectivity profile of a new class of ligands, 1,4-acylthiazepanes, and find them to have ≥3-10-fold selectivity for the C-terminal bromodomain of both BRD4 and BRDT. Given the speed and lower protein concentration required over traditional protein-observed NMR methods, we envision that these fluorinated tandem proteins may find use in fragment screening and evaluating nucleosome and transcription factor interactions.