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

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

doi:10.3390/ijms23063184

Open AccessArticle

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

by

Eléonore W. E. Verweij

^†,

Reggie Bosma

^†,

Meichun Gao

,

Jelle van den Bor

,

Betty Al Araaj

,

Sabrina M. de Munnik

,

Xiaoyuan Ma

,

Rob Leurs

and

Henry F. Vischer

^*

Division of Medicinal Chemistry, Faculty of Science, Amsterdam Institute of Molecular and Life Sciences, Vrije Universiteit Amsterdam, 1081 HZ Amsterdam, The Netherlands

^*

Author to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Int. J. Mol. Sci. 2022, 23(6), 3184; https://doi.org/10.3390/ijms23063184

Submission received: 21 February 2022 / Revised: 10 March 2022 / Accepted: 11 March 2022 / Published: 16 March 2022

(This article belongs to the Special Issue Biased Agonism in GPCRs: An Opportunity for Drug Discovery)

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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 G_q 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 G_q 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 G_q biosensor activation.

Keywords: histamine; H₁R; GPCR; G protein; β-arrestin; GPCR kinase; BRET assay; biased signaling

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MDPI and ACS Style

Verweij, E.W.E.; Bosma, R.; Gao, M.; van den Bor, J.; Al Araaj, B.; de Munnik, S.M.; Ma, X.; Leurs, R.; Vischer, H.F. BRET-Based Biosensors to Measure Agonist Efficacies in Histamine H₁ Receptor-Mediated G Protein Activation, Signaling and Interactions with GRKs and β-Arrestins. Int. J. Mol. Sci. 2022, 23, 3184. https://doi.org/10.3390/ijms23063184

AMA Style

Verweij EWE, Bosma R, Gao M, van den Bor J, Al Araaj B, de Munnik SM, Ma X, Leurs R, Vischer HF. BRET-Based Biosensors to Measure Agonist Efficacies in Histamine H₁ Receptor-Mediated G Protein Activation, Signaling and Interactions with GRKs and β-Arrestins. International Journal of Molecular Sciences. 2022; 23(6):3184. https://doi.org/10.3390/ijms23063184

Chicago/Turabian Style

Verweij, Eléonore W. E., Reggie Bosma, Meichun Gao, Jelle van den Bor, Betty Al Araaj, Sabrina M. de Munnik, Xiaoyuan Ma, Rob Leurs, and Henry F. Vischer. 2022. "BRET-Based Biosensors to Measure Agonist Efficacies in Histamine H₁ Receptor-Mediated G Protein Activation, Signaling and Interactions with GRKs and β-Arrestins" International Journal of Molecular Sciences 23, no. 6: 3184. https://doi.org/10.3390/ijms23063184

APA Style

Verweij, E. W. E., Bosma, R., Gao, M., van den Bor, J., Al Araaj, B., de Munnik, S. M., Ma, X., Leurs, R., & Vischer, H. F. (2022). BRET-Based Biosensors to Measure Agonist Efficacies in Histamine H₁ Receptor-Mediated G Protein Activation, Signaling and Interactions with GRKs and β-Arrestins. International Journal of Molecular Sciences, 23(6), 3184. https://doi.org/10.3390/ijms23063184

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

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