The DOI matches its Crossref record; 59 of the 60 references it cites were found in a registry or an index.
High-Throughput Screening of G Protein-Coupled Receptor Antagonists Using a Bioluminescence Resonance Energy Transfer 1-Based β-Arrestin2 Recruitment Assay
Open accessAbstract
In this study, the authors developed HEK293 cell lines that stably coexpressed optimal amounts of beta-arrestin2-Rluc and VENUS fusions of G protein-coupled receptors (GPCRs) belonging to both class A and class B receptors, which include receptors that interact transiently or stably with beta-arrestins. This allowed the use of a bioluminescence resonance energy transfer (BRET) 1- beta-arrestin2 translocation assay to quantify receptor activation or inhibition. One of the developed cell lines coexpressing CCR5-VENUS and beta-arrestin2- Renilla luciferase was then used for high-throughput screening (HTS) for antagonists of the chemokine receptor CCR5, the primary co-receptor for HIV. A total of 26,000 compounds were screened for inhibition of the agonist-promoted beta-arrestin2 recruitment to CCR5, and 12 compounds were found to specifically inhibit the agonist-induced beta-arrestin2 recruitment to CCR5. Three of the potential hits were further tested using other functional assays, and their abilities to inhibit CCR5 agonist-promoted signaling were confirmed. This is the 1st study describing a BRET1-beta-arrestin recruitment assay in stable mammalian cells and its successful application in HTS for GPCRs antagonists.
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High-throughput screening assays for the identification of chemical probes
Inglese, J., Johnson, R. L., Simeonov, A. M., Xia, M., Zheng, W., Austin, C. P., & Auld, D. S. (2007) · Nature Chemical Biology · doi:10.1038/nchembio.2007.17 · Cited by 632
Kenakin, T., & Miller, L. J. (2010) · Pharmacological Reviews · doi:10.1124/pr.108.000992 · Cited by 584
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NanoBRET─A Novel BRET Platform for the Analysis of Protein–Protein Interactions
Machleidt, T., Woodroofe, C. C., Schwinn, M. K., Méndez, J., Robers, M. B., Zimmerman, K., Otto, P., Daniels, D. L., Kirkland, T. A., & Wood, K. V. (2015) · ACS Chemical Biology · doi:10.1021/acschembio.5b00143 · Cited by 504
Modulators of Protein–Protein Interactions
Milroy, L.-G., Grossmann, T. N., Hennig, S., Brunsveld, L., & Ottmann, C. (2014) · Chemical Reviews · doi:10.1021/cr400698c · Cited by 497
Metadata
- Authors
- Hamdan, F. F., Audet, M., Garneau, P., Pelletier, J., & Bouvier, M.
- Affiliations
- Université de Montréal (CA)
- McGill University (CA)
- Venue
- SLAS Discovery
- Publisher
- Elsevier BV
- Published
- 2005-08
- Volume
- 10
- Issue
- 5
- Pages
- 463-475
- ISSN
- 2472-5552 (ISSN-L)
- OpenAlex
- W2094714066
- Cited by
- 183
- References
- 60
- Citations by year
- 2026: 2 · 2025: 4 · 2024: 2 · 2023: 3 · 2022: 4 · 2021: 9 · 2020: 8 · 2019: 6 · 2018: 8 · 2017: 5 · 2016: 6 · 2015: 17 · 2014: 8 · 2013: 8 · 2012: 16
- FWCI
- 4.64
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- SLAS DISCOVERY (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › bioluminescence and chemiluminescence research
- Health Sciences › Medicine › Oncology › Chemokine receptors and signaling
- Keywords
- G protein-coupled receptors, β-arrestin2, CCR5, chemokine receptor antagonists, high-throughput screening, HIV co-receptor, HEK293 cells, receptor antagonists
- MeSH terms
- beta-Arrestins, Animals, Automation, Calcium, Cell Line, Dose-Response Relationship, Drug, Drug Evaluation, Preclinical, Energy Transfer, HIV, Humans, Luminescent Measurements, Microscopy, Fluorescence, Plasmids, Time Factors, Genes, Reporter, Arrestins, Receptors, CCR5, Protein Transport, Receptors, G-Protein-Coupled, Macromolecular Substances, Renilla, Luciferases, Renilla, Green Fluorescent Proteins
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