A NanoBRET-Based H3R Conformational Biosensor to Study Real-Time H3 Receptor Pharmacology in Cell Membranes and Living Cells

Xiaoyuan Ma, Meichun Gao, Henry F. Vischer, Rob Leurs

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Conformational biosensors to monitor the activation state of G protein-coupled receptors are a useful addition to the molecular pharmacology assay toolbox to characterize ligand efficacy at the level of receptor proteins instead of downstream signaling. We recently reported the initial characterization of a NanoBRET-based conformational histamine H3 receptor (H3R) biosensor that allowed the detection of both (partial) agonism and inverse agonism on living cells in a microplate reader assay format upon stimulation with H3R ligands. In the current study, we have further characterized this H3R biosensor on intact cells by monitoring the effect of consecutive ligand injections in time and evaluating its compatibility with photopharmacological ligands that contain a light-sensitive azobenzene moiety for photo-switching. In addition, we have validated the H3R biosensor in membrane preparations and found that observed potency values better correlated with binding affinity values that were measured in radioligand competition binding assays on membranes. Hence, the H3R conformational biosensor in membranes might be a ready-to-use, high-throughput alternative for radioligand binding assays that in addition can also detect ligand efficacies with comparable values as the intact cell assay.

Original languageEnglish
Article number8211
Pages (from-to)1-13
Number of pages13
JournalInternational Journal of Molecular Sciences
Volume23
Issue number15
Early online date26 Jul 2022
DOIs
Publication statusPublished - 1 Aug 2022

Bibliographical note

Special Issue: Molecular Biology of Histamine Systems 2.0.

Funding

X.M. (201703250074) and M.G. (202006310027) are supported by CSC Chinese scholarship grants.

FundersFunder number
China Scholarship Council

    Keywords

    • BRET
    • conformational biosensor
    • GPCR
    • H3R
    • histamine

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