Direct NMR detection of the binding of functional ligands to the human sweet receptor, a heterodimeric family 3 GPCR

Fariba M. Assadi-Porter, Marco Tonelli, Emeline Maillet, Klaas Hallenga, Outhiriaradjou Benard, Marianna Max, John L. Markley

Research output: Contribution to journalArticle

48 Scopus citations

Abstract

We present a robust method for monitoring the binding of ligands to the heterodimeric (T1R2+T1R3) human sweet receptor (a family 3 GPCR receptor). The approach utilizes saturation transfer difference (STD) NMR spectroscopy with receptor proteins expressed on the surface of human epithelial kidney cells. The preparation investigated by NMR can contain either live cells or membranes isolated from these cells containing the receptor. We have used this approach to confirm the noncompetitive binding of alitame and cyclamate to the receptor and to determine that greatly reduced receptor binding affinity compared to wild-type brazzein explains the lack of sweetness of brazzein mutant A16C17. This approach opens new avenues for research on the mechanism of action of the sweet receptor and for the design of new noncalorigenic sweeteners.

Original languageEnglish (US)
Pages (from-to)7212-7213
Number of pages2
JournalJournal of the American Chemical Society
Volume130
Issue number23
DOIs
StatePublished - Jun 11 2008

ASJC Scopus subject areas

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

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