TY - JOUR
T1 - Quantifying the selectivity of protein−protein and small molecule interactions with fluorinated tandem bromodomain reader proteins
AU - Pomerantz, William C.K.
AU - Kalra, Prakriti
AU - McGraw, Logan
AU - Kimbrough, Jennifer R.
AU - Pandey, Anil K.
AU - Solberg, Jonathan
AU - Cui, Huarui
AU - Divakaran, Anand
AU - John, Kristen
AU - Hawkinson, Jon E.
N1 - Publisher Copyright:
© 2020 American Chemical Society
PY - 2020/11/20
Y1 - 2020/11/20
N2 - 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.
AB - 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.
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U2 - 10.1021/acschembio.0c00720
DO - 10.1021/acschembio.0c00720
M3 - Article
C2 - 33138352
AN - SCOPUS:85096508791
SN - 1554-8929
VL - 15
SP - 3038
EP - 3049
JO - ACS Chemical Biology
JF - ACS Chemical Biology
IS - 11
ER -