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 1H,15N HSQC/HMQC NMR spectroscopy. Metabolic labeling with multiple fluorinated amino acids provides useful probes for further studying ligand binding and has led to new insight for allosterically regulating transcription-factor protein interactions with small-molecule ligands.
Original language | English (US) |
---|---|
Pages (from-to) | 963-969 |
Number of pages | 7 |
Journal | ChemBioChem |
Volume | 19 |
Issue number | 9 |
DOIs | |
State | Published - May 4 2018 |
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Keywords
- NMR spectroscopy
- allosterism
- biomolecular NMR
- fluorine
- protein–protein interactions
PubMed: MeSH publication types
- Journal Article
- Research Support, N.I.H., Extramural
- Research Support, Non-U.S. Gov't
- Research Support, U.S. Gov't, Non-P.H.S.
Cite this
Dual Labeling of the CBP/p300 KIX Domain for 19F NMR Leads to Identification of a New Small-Molecule Binding Site. / Gee, Clifford T.; Arntson, Keith E.; Koleski, Edward J.; Staebell, Rachel Lynn; Pomerantz, William C.
In: ChemBioChem, Vol. 19, No. 9, 04.05.2018, p. 963-969.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Dual Labeling of the CBP/p300 KIX Domain for 19F NMR Leads to Identification of a New Small-Molecule Binding Site
AU - Gee, Clifford T.
AU - Arntson, Keith E.
AU - Koleski, Edward J.
AU - Staebell, Rachel Lynn
AU - Pomerantz, William C
PY - 2018/5/4
Y1 - 2018/5/4
N2 - 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 1H,15N HSQC/HMQC NMR spectroscopy. Metabolic labeling with multiple fluorinated amino acids provides useful probes for further studying ligand binding and has led to new insight for allosterically regulating transcription-factor protein interactions with small-molecule ligands.
AB - 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 1H,15N HSQC/HMQC NMR spectroscopy. Metabolic labeling with multiple fluorinated amino acids provides useful probes for further studying ligand binding and has led to new insight for allosterically regulating transcription-factor protein interactions with small-molecule ligands.
KW - NMR spectroscopy
KW - allosterism
KW - biomolecular NMR
KW - fluorine
KW - protein–protein interactions
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UR - http://www.scopus.com/inward/citedby.url?scp=85044852918&partnerID=8YFLogxK
U2 - 10.1002/cbic.201700686
DO - 10.1002/cbic.201700686
M3 - Article
C2 - 29430847
AN - SCOPUS:85044852918
VL - 19
SP - 963
EP - 969
JO - ChemBioChem
JF - ChemBioChem
SN - 1439-4227
IS - 9
ER -