BRET-based beta-arrestin2 recruitment to the histamine H-1 receptor for investigating antihistamine binding kinetics

作者：Bosma Reggie; Moritani Ryo; Leurs Rob; Vischer Henry F

来源：Pharmacological Research, 2016, 111: 679-687.

DOI：10.1016/j.phrs.2016.07.034

• Histamine H-1 receptor
• Arrestin
• Binding rate
• Residence time
• Bioluminescence resonance energy-transfer
• BRET

Ligand residence time is thought to be a critical parameter for optimizing the in vivo efficacy of drug candidates. For the histamine H-1 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 beta-arresting 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 beta-arresting to the H1R and the transiently recruited beta-arrestin2 therefore reflected occupancy of the receptor by histamine. Antihistamines displayed similar kinetic signatures on antagonizing histamine-induced beta-arrestin-2 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.

• 出版日期2016-9