Quantifying the selectivity of protein−protein and small molecule interactions with fluorinated tandem bromodomain reader proteins

William C.K. Pomerantz, Prakriti Kalra, Logan McGraw, Jennifer R. Kimbrough, Anil K. Pandey, Jonathan Solberg, Huarui Cui, Anand Divakaran, Kristen John, Jon E. Hawkinson

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish (US)
Pages (from-to)3038-3049
Number of pages12
JournalACS Chemical Biology
Volume15
Issue number11
DOIs
StatePublished - Nov 20 2020

Bibliographical note

Funding Information:
Financial support for this project is acknowledged from the Masonic Cancer Center at the University of Minnesota and the NICHD (HHSN27520130001 and 1 U54 HD09354).

Publisher Copyright:
© 2020 American Chemical Society

PubMed: MeSH publication types

  • Journal Article
  • Research Support, N.I.H., Extramural

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