title: BRET-based β-arrestin2 recruitment to the histamine H1 receptor for investigating antihistamine binding kinetics.
abstract: 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.
authors.affiliation: Department of Chemistry and Pharmaceutical Sciences, Division of Medicinal Chemistry, Amsterdam Institute for Molecules, Medicines and Systems (AIMMS), Vrije Universiteit Amsterdam, PO Box 7161, 1007 MC Amsterdam, The Netherlands.
authors.forename: Reggie
authors.initials: R
authors.lastname: Bosma
authors.affiliation: Department of Chemistry and Pharmaceutical Sciences, Division of Medicinal Chemistry, Amsterdam Institute for Molecules, Medicines and Systems (AIMMS), Vrije Universiteit Amsterdam, PO Box 7161, 1007 MC Amsterdam, The Netherlands.
authors.forename: Ryo
authors.initials: R
authors.lastname: Moritani
authors.affiliation: Department of Chemistry and Pharmaceutical Sciences, Division of Medicinal Chemistry, Amsterdam Institute for Molecules, Medicines and Systems (AIMMS), Vrije Universiteit Amsterdam, PO Box 7161, 1007 MC Amsterdam, The Netherlands.
authors.forename: Rob
authors.initials: R
authors.lastname: Leurs
authors.affiliation: Department of Chemistry and Pharmaceutical Sciences, Division of Medicinal Chemistry, Amsterdam Institute for Molecules, Medicines and Systems (AIMMS), Vrije Universiteit Amsterdam, PO Box 7161, 1007 MC Amsterdam, The Netherlands. Electronic address: h.f.vischer@vu.nl.
authors.forename: Henry F
authors.initials: HF
authors.lastname: Vischer
mesh_terms: D001667:Binding, Competitive; D060807:Bioluminescence Resonance Energy Transfer Techniques; D017332:Cetirizine / Q000378:metabolism* / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D057809:HEK293 Cells; D006632:Histamine / Q000378:metabolism / Q000494:pharmacology; D017442:Histamine Agonists / Q000378:metabolism / Q000494:pharmacology; D039563:Histamine H1 Antagonists, Non-Sedating / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008024:Ligands; D017336:Loratadine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D049409:Luciferases, Firefly / Q000235:genetics / Q000378:metabolism; D008954:Models, Biological; D050778:NFATC Transcription Factors / Q000235:genetics; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011869:Radioligand Assay; D011969:Receptors, Histamine H1 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D000071560:beta-Arrestin 2 / Q000378:metabolism*
chemical_list: C000605254:ARRB2 protein, human; D017442:Histamine Agonists; D039563:Histamine H1 Antagonists, Non-Sedating; D008024:Ligands; D050778:NFATC Transcription Factors; D011969:Receptors, Histamine H1; D011993:Recombinant Fusion Proteins; D000071560:beta-Arrestin 2; C472067:levocetirizine; D017336:Loratadine; D006632:Histamine; D049409:Luciferases, Firefly; C121345:desloratadine; D017332:Cetirizine
keywords: Arrestin; BRET; Binding rate; Bioluminescence resonance energy-transfer; Histamine H(1) receptor; Residence time