Time-resolved FRET between GPCR ligands reveals oligomers in native tissues

Nat Chem Biol. 2010 Aug;6(8):587-94. doi: 10.1038/nchembio.396. Epub 2010 Jul 11.


G protein-coupled receptor (GPCR) oligomers have been proposed to play critical roles in cell signaling, but confirmation of their existence in a native context remains elusive, as no direct interactions between receptors have been reported. To demonstrate their presence in native tissues, we developed a time-resolved FRET strategy that is based on receptor labeling with selective fluorescent ligands. Specific FRET signals were observed with four different receptors expressed in cell lines, consistent with their dimeric or oligomeric nature in these transfected cells. More notably, the comparison between FRET signals measured with sets of fluorescent agonists and antagonists was consistent with an asymmetric relationship of the two protomers in an activated GPCR dimer. Finally, we applied the strategy to native tissues and succeeded in demonstrating the presence of oxytocin receptor dimers and/or oligomers in mammary gland.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Animals
  • Antidiuretic Hormone Receptor Antagonists
  • COS Cells
  • Cell Line
  • Chlorocebus aethiops
  • Dimerization
  • Dopamine D2 Receptor Antagonists
  • Female
  • Fluorescence Resonance Energy Transfer / methods*
  • Fluorescent Dyes
  • Ligands
  • Mammary Glands, Animal / metabolism
  • Models, Molecular
  • Oligopeptides / chemistry*
  • Oligopeptides / metabolism
  • Radioligand Assay
  • Rats
  • Receptors, Dopamine D2 / agonists
  • Receptors, Dopamine D2 / metabolism
  • Receptors, G-Protein-Coupled / agonists
  • Receptors, G-Protein-Coupled / antagonists & inhibitors
  • Receptors, G-Protein-Coupled / metabolism*
  • Receptors, Oxytocin / agonists
  • Receptors, Oxytocin / antagonists & inhibitors
  • Receptors, Oxytocin / metabolism
  • Receptors, Vasopressin / agonists
  • Receptors, Vasopressin / metabolism


  • Antidiuretic Hormone Receptor Antagonists
  • Dopamine D2 Receptor Antagonists
  • Fluorescent Dyes
  • Ligands
  • Oligopeptides
  • Receptors, Dopamine D2
  • Receptors, G-Protein-Coupled
  • Receptors, Oxytocin
  • Receptors, Vasopressin

Associated data

  • PubChem-Substance/96021130
  • PubChem-Substance/96021131