Competition saturation transfer difference experiments improved with isotope editing and filtering schemes in NMR-based screening

Krisztina Fehér, Patrick Groves, G. Batta, Jesús Jiménez-Barbero, Claudia Muhle-Goll, K. Kövér

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Competition binding experiments are used in NMR-based screenings to match up to the binding site with that of a known ligand and to determine the strength of the interaction, thus providing a ranking of hits according to receptor affinity. These competition titration experiments must use a reference ligand for which the binding site on the receptor and the affinity of the interaction is known. These experiments rely on the observation of separate signals of the reference and hit compounds, which is very often hampered by signal overlap. Here, we present a combination of isotope editing and filtering schemes with saturation transfer difference (STD) experiments that allows the separation of the STD signals of the labeled reference ligand from that of the natural abundance hit compound even in the case of severe signal overlap. Thus, the measurement of well-defined titration curves in competition STD titration experiments is feasible and allows the quantitative determination of binding constants. Note that the method requires the availability of the reference ligand in an NMR-active, stable-isotope-labeled form.

Original languageEnglish
Pages (from-to)17148-17153
Number of pages6
JournalJournal of the American Chemical Society
Volume130
Issue number50
DOIs
Publication statusPublished - Dec 17 2008

Fingerprint

Isotopes
Screening
Nuclear magnetic resonance
Ligands
Titration
Binding sites
Experiments
Binding Sites
Observation
Availability

ASJC Scopus subject areas

  • Chemistry(all)
  • Catalysis
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Competition saturation transfer difference experiments improved with isotope editing and filtering schemes in NMR-based screening. / Fehér, Krisztina; Groves, Patrick; Batta, G.; Jiménez-Barbero, Jesús; Muhle-Goll, Claudia; Kövér, K.

In: Journal of the American Chemical Society, Vol. 130, No. 50, 17.12.2008, p. 17148-17153.

Research output: Contribution to journalArticle

Fehér, Krisztina ; Groves, Patrick ; Batta, G. ; Jiménez-Barbero, Jesús ; Muhle-Goll, Claudia ; Kövér, K. / Competition saturation transfer difference experiments improved with isotope editing and filtering schemes in NMR-based screening. In: Journal of the American Chemical Society. 2008 ; Vol. 130, No. 50. pp. 17148-17153.
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