BRET-based β-arrestin2 recruitment to the histamine H1 receptor for investigating antihistamine binding kinetics

Pharmacol Res. 2016 Sep;111:679-687. doi: 10.1016/j.phrs.2016.07.034. Epub 2016 Jul 26.


Ligand residence time is thought to be a critical parameter for optimizing the in vivo efficacy of drug candidates. For the histamine H1 receptor (H1R) and other G protein-coupled receptors, the kinetics of ligand binding are typically measured by low throughput radioligand binding experiments using homogenized cell membranes expressing the target receptor. In this study, a real-time proximity assay between H1R and β-arrestin2 in living cells was established to investigate the dynamics of antihistamine binding to the H1R. No receptor reserve was found for the histamine-induced recruitment of β-arrestin2 to the H1R and the transiently recruited β-arrestin2 therefore reflected occupancy of the receptor by histamine. Antihistamines displayed similar kinetic signatures on antagonizing histamine-induced β-arrestin2 recruitment as compared to displacing radioligand binding from the H1R. This homogeneous functional method unambiguously determined the fifty-fold difference in the dissociation rate constant between mepyramine and the long residence time antihistamines levocetirizine and desloratadine.

Keywords: Arrestin; BRET; Binding rate; Bioluminescence resonance energy-transfer; Histamine H(1) receptor; Residence time.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding, Competitive
  • Bioluminescence Resonance Energy Transfer Techniques*
  • Cetirizine / metabolism*
  • Cetirizine / pharmacology
  • Dose-Response Relationship, Drug
  • HEK293 Cells
  • Histamine / metabolism
  • Histamine / pharmacology
  • Histamine Agonists / metabolism
  • Histamine Agonists / pharmacology
  • Histamine H1 Antagonists, Non-Sedating / metabolism*
  • Histamine H1 Antagonists, Non-Sedating / pharmacology
  • Humans
  • Kinetics
  • Ligands
  • Loratadine / analogs & derivatives*
  • Loratadine / metabolism
  • Loratadine / pharmacology
  • Luciferases, Firefly / genetics
  • Luciferases, Firefly / metabolism
  • Models, Biological
  • NFATC Transcription Factors / genetics
  • Promoter Regions, Genetic
  • Protein Binding
  • Radioligand Assay
  • Receptors, Histamine H1 / drug effects
  • Receptors, Histamine H1 / genetics
  • Receptors, Histamine H1 / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • beta-Arrestin 2 / metabolism*


  • ARRB2 protein, human
  • Histamine Agonists
  • Histamine H1 Antagonists, Non-Sedating
  • Ligands
  • NFATC Transcription Factors
  • Receptors, Histamine H1
  • Recombinant Fusion Proteins
  • beta-Arrestin 2
  • levocetirizine
  • Loratadine
  • Histamine
  • Luciferases, Firefly
  • desloratadine
  • Cetirizine