Monitoring G protein-coupled receptor activation using an adenovirus-based ß-arrestin bimolecular fluorescence complementation assay.
Anal Biochem
; 449: 32-41, 2014 Mar 15.
Article
em En
| MEDLINE
| ID: mdl-24361713
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 ß-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 ß-arrestin BiFC assay in cells transduced with recombinant adenoviruses. An analysis of the activation of somatostatin receptor 2 (SSTR2) with the adenovirus-based ß-arrestin BiFC assay showed that the assay is suitable for quantifying SSTR2 activation in response to specific agonists or antagonists. Furthermore, the adenovirus-based ß-arrestin BiFC assay was able to detect the activation of a broad range of GPCRs. Collectively, our data indicate that the adenovirus-based ß-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.
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Base de dados:
MEDLINE
Assunto principal:
Técnicas Biossensoriais
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Adenoviridae
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Arrestinas
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Receptores Acoplados a Proteínas G
Limite:
Humans
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article