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Detection and quantification of allosteric modulation of endogenous M4 muscarinic acetylcholine receptor using impedance-based label-free technology in a neuronal cell line

Chen, Amy N.Y.; Malone, Daniel T.; Pabreja, Kavita; Sexton, Patrick M.; Christopoulos, Arthur; Canals, Meritxell

Authors

Amy N.Y. Chen

Daniel T. Malone

Kavita Pabreja

Patrick M. Sexton

Arthur Christopoulos



Abstract

© 2014 Society for Laboratory Automation and Screening. Allosteric modulators of G protein-coupled receptors have the potential to achieve greater receptor subtype selectivity compared with ligands targeting the orthosteric site of this receptor family. However, the high attrition rate in GPCR drug discovery programs has highlighted the need to better characterize lead compounds in terms of their allosteric action, as well as the signals they elicit. Recently, the use of label-free technologies has been proposed as an approach to overcome some limitations of endpoint-based assays and detect global changes in the ligand-stimulated cell. In this study, we assessed the ability of an impedance-based label-free technology, xCELLigence, to detect allosteric modulation in a neuronal cell line natively expressing rodent M4 muscarinic acetylcholine receptors. We were able to demonstrate that positive allosteric modulation of the endogenous M4 muscarinic acetylcholine receptor can be detected using this technology. Importantly, the allosteric parameters estimated from the label-free approach are comparable to those estimated from endpoint-based assays.

Journal Article Type Article
Publication Date Dec 22, 2014
Journal Journal of Biomolecular Screening
Print ISSN 1087-0571
Electronic ISSN 1552-454X
Publisher SAGE Publications
Peer Reviewed Peer Reviewed
Volume 20
Issue 5
Pages 646-654
APA6 Citation Chen, A. N., Chen, A., Malone, D. T., Pabreja, K., Sexton, P. M., Christopoulos, A., & Canals, M. (2014). Detection and quantification of allosteric modulation of endogenous M4 muscarinic acetylcholine receptor using impedance-based label-free technology in a neuronal cell line. Journal of Biomolecular Screening, 20(5), 646-654. https://doi.org/10.1177/1087057114563025
DOI https://doi.org/10.1177/1087057114563025
Keywords cell impedance; G protein-coupled receptor; label-free technology; muscarinic acetylcholine receptor; positive allosteric modulator; acetylcholine; agents interacting with transmitter, hormone or drug receptors; ly 2033298; muscarinic M4 receptor; unclass
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