Monitoring G protein-coupled receptor activation using an adenovirus-based beta-arrestin bimolecular fluorescence complementation assay
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Title
- Monitoring G protein-coupled receptor activation using an adenovirus-based beta-arrestin bimolecular fluorescence complementation assay
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Author(s)
- Song, Yong Bhum; Park, Chul O.; Jeong, Jae-Yeon; Huh, Won-Ki
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Alternative Author(s)
- 정재연
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Publication Year
- 2014-03-15
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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 beta-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 beta-arrestin BiFC assay in cells transduced with recombinant adenoviruses. An analysis of the activation of somatostatin receptor 2 (SSTR2) with the adenovirus-based beta-arrestin BiFC assay showed that the assay is suitable for quantifying SSTR2 activation in response to specific agonists or antagonists. Furthermore, the adenovirus-based beta-arrestin BiFC assay was able to detect the activation of a broad range of GPCRs. Collectively, our data indicate that the adenovirus-based beta-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. (C) 2013 Elsevier Inc. All rights reserved.
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ISSN
- 0003-2697
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URI
- https://sciwatch.kiost.ac.kr/handle/2020.kiost/2891
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DOI
- 10.1016/j.ab.2013.12.017
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Bibliographic Citation
- ANALYTICAL BIOCHEMISTRY, v.449, pp.32 - 41, 2014
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Publisher
- ACADEMIC PRESS INC ELSEVIER SCIENCE
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Subject
- DRUG DISCOVERY; 7-TRANSMEMBRANE RECEPTORS; CELLS; BETA-ARRESTIN2; VISUALIZATION; TRAFFICKING; DEGRADATION; ENDOCYTOSIS; COMPLEXES
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Keywords
- G protein-coupled receptor (GPCR); beta-Arrestin; Bimolecular fluorescence complementation (BiFC); Adenovirus; High-throughput screening
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Type
- Article
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Language
- English
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Document Type
- Article
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