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Monitoring G protein-coupled receptor activation using an adenovirus-based β-arrestin bimolecular fluorescence

Yong Bhum Song1, Chul O Park1, Jae-Yeon Jeong2

  • 1Department of Biological Sciences and Research Center for Functional Cellulomics, Seoul National University, Seoul 151-747, Republic of Korea.

Analytical Biochemistry
|December 24, 2013
PubMed

Insights

This study introduces a new, high-throughput adenovirus-based assay for studying G protein-coupled receptors (GPCRs). This method simplifies the analysis of GPCR activation and drug screening for various diseases.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Pharmacology

Background:

  • G protein-coupled receptors (GPCRs) are crucial cell-surface receptors implicated in numerous diseases.
  • GPCRs are significant therapeutic targets, necessitating efficient methods for studying their activation.
  • The beta-arrestin recruitment assay is a key tool for analyzing GPCR activation.

Purpose of the Study:

  • To develop a high-throughput method for cloning GPCR cDNAs into adenoviral vectors.
  • To establish and validate an adenovirus-based beta-arrestin bimolecular fluorescence complementation (BiFC) assay for GPCR activation.
  • To demonstrate the utility of this assay for drug screening targeting GPCRs.

Main Methods:

  • Cloning of GPCR cDNAs into adenoviral BiFC vectors.
  • Transduction of cells with recombinant adenoviruses.
  • Performing beta-arrestin BiFC assays to quantify GPCR activation.

Main Results:

  • The adenovirus-based beta-arrestin BiFC assay successfully quantified somatostatin receptor 2 (SSTR2) activation by agonists and antagonists.
  • The assay demonstrated broad applicability, detecting activation across a wide range of GPCRs.
  • The method proved suitable for high-throughput analysis.

Conclusions:

  • The adenovirus-based beta-arrestin BiFC assay provides a simple and universal platform for studying GPCR activation.
  • This assay is valuable for high-throughput screening of drugs targeting GPCRs.
  • The developed method facilitates research into GPCRs and associated diseases.
Keywords:
AdenovirusBimolecular fluorescence complementation (BiFC)G protein-coupled receptor (GPCR)High-throughput screeningβ-Arrestin

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