BRET-Based Biosensors to Measure Agonist Efficacies in Histamine H1 Receptor-Mediated G Protein Activation, Signaling and Interactions with GRKs and β-Arrestins

Eléonore W.E. Verweij, Reggie Bosma, Meichun Gao, Jelle van den Bor, Betty Al Araaj, Sabrina M. de Munnik, Xiaoyuan Ma, Rob Leurs, Henry F. Vischer*

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

The histamine H1 receptor (H1R) is a G protein-coupled receptor (GPCR) and plays a key role in allergic reactions upon activation by histamine which is locally released from mast cells and basophils. Consequently, H1R is a well-established therapeutic target for antihistamines that relieve allergy symptoms. H1R signals via heterotrimeric Gq proteins and is phosphorylated by GPCR kinase (GRK) subtypes 2, 5, and 6, consequently facilitating the subsequent recruitment of β-arrestin1 and/or 2. Stimulation of a GPCR with structurally different agonists can result in preferential engagement of one or more of these intracellular signaling molecules. To evaluate this so-called biased agonism for H1R, bioluminescence resonance energy transfer (BRET)-based biosensors were applied to measure H1R signaling through heterotrimeric Gq proteins, second messengers (inositol 1,4,5-triphosphate and Ca2+), and receptor-protein interactions (GRKs and β-arrestins) in response to histamine, 2-phenylhistamines, and histaprodifens in a similar cellular background. Although differences in efficacy were observed for these agonists between some functional readouts as compared to reference agonist histamine, subsequent data analysis using an operational model of agonism revealed only signaling bias of the agonist Br-phHA-HA in recruiting β-arrestin2 to H1R over Gq biosensor activation.

Original languageEnglish
Article number3184
Pages (from-to)1-17
Number of pages17
JournalInternational Journal of Molecular Sciences
Volume23
Issue number6
DOIs
Publication statusPublished - 2 Mar 2022

Bibliographical note

Funding Information:
Funding: This research was supported by The Netherlands Organisation for Scientific Research (NWO) ECHO project 711.013.014 (R.L. and H.F.V.), EU/EFPIA Innovative Medicines Initiative (IMI) Joint Undertaking, K4DD (grant 115366). X.M. (201703250074) and M.G. (202006310027) are supported by CSC Chinese scholarship grants.

Publisher Copyright:
© 2022 by the authors. Licensee MDPI, Basel, Switzerland.

Keywords

  • Biased signaling
  • BRET assay
  • G protein
  • GPCR
  • GPCR kinase
  • HR
  • Histamine
  • β-arrestin

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