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

Clifford T. Gee, Keith E. Arntson, Edward J. Koleski, Rachel Lynn Staebell, William C Pomerantz

Research output: Contribution to journalArticle

3 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)963-969
Number of pages7
JournalChemBioChem
Volume19
Issue number9
DOIs
StatePublished - May 4 2018

Fingerprint

Labeling
Binding Sites
Nuclear magnetic resonance
Ligands
Molecules
Fluorine
Amino Acids
Nuclear magnetic resonance spectroscopy
Proteins
Transcription Factors
Magnetic Resonance Spectroscopy
Labels
Screening

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 journalArticle

Gee, Clifford T. ; Arntson, Keith E. ; Koleski, Edward J. ; Staebell, Rachel Lynn ; Pomerantz, William C. / Dual Labeling of the CBP/p300 KIX Domain for 19F NMR Leads to Identification of a New Small-Molecule Binding Site. In: ChemBioChem. 2018 ; Vol. 19, No. 9. pp. 963-969.
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