title: Monitoring G protein-coupled receptor activation using an adenovirus-based β-arrestin bimolecular fluorescence complementation assay.
abstract: G protein-coupled receptors (GPCRs) are the largest family of cell-surface receptors and are involved in a variety of pathological conditions including cancer and cardiovascular, metabolic, neurological, and autoimmune diseases. GPCRs are being intensively investigated as targets for therapeutic intervention, and the β-arrestin recruitment assay has become a popular tool for analyzing GPCR activation. Here, we report a high-throughput method for cloning GPCR cDNAs into adenoviral bimolecular fluorescence complementation (BiFC) vectors and performing the β-arrestin BiFC assay in cells transduced with recombinant adenoviruses. An analysis of the activation of somatostatin receptor 2 (SSTR2) with the adenovirus-based β-arrestin BiFC assay showed that the assay is suitable for quantifying SSTR2 activation in response to specific agonists or antagonists. Furthermore, the adenovirus-based β-arrestin BiFC assay was able to detect the activation of a broad range of GPCRs. Collectively, our data indicate that the adenovirus-based β-arrestin BiFC assay can serve as a simple and universal platform for studying GPCR activation and thus will be useful for high-throughput screening of drugs that target GPCRs.
authors.affiliation: Department of Biological Sciences and Research Center for Functional Cellulomics, Seoul National University, Seoul 151-747, Republic of Korea.
authors.forename: Yong Bhum
authors.initials: YB
authors.lastname: Song
authors.affiliation: Department of Biological Sciences and Research Center for Functional Cellulomics, Seoul National University, Seoul 151-747, Republic of Korea.
authors.forename: Chul O
authors.initials: CO
authors.lastname: Park
authors.affiliation: Marine Biotechnology Research Division, Korea Institute of Ocean Science & Technology, Ansan 426-744, Republic of Korea. Electronic address: jeongjy@kiost.ac.
authors.forename: Jae-Yeon
authors.initials: JY
authors.lastname: Jeong
authors.affiliation: Department of Biological Sciences and Research Center for Functional Cellulomics, Seoul National University, Seoul 151-747, Republic of Korea; Institute of Microbiology, Seoul National University, Seoul 151-747, Republic of Korea. Electronic address: wkh@snu.ac.kr.
authors.forename: Won-Ki
authors.initials: WK
authors.lastname: Huh
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D019390:Arrestins / Q000378:metabolism*; D015374:Biosensing Techniques / Q000379:methods*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005453:Fluorescence; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D000071557:beta-Arrestins
chemical_list: D019390:Arrestins; D018076:DNA, Complementary; D043562:Receptors, G-Protein-Coupled; D000071557:beta-Arrestins
keywords: Adenovirus; Bimolecular fluorescence complementation (BiFC); G protein-coupled receptor (GPCR); High-throughput screening; β-Arrestin