Ngo, Tony and Coleman, James L J and Smith, Nicola J (2015) Using constitutive activity to define appropriate high-throughput screening assays for orphan G protein-coupled receptors. Methods in Molecular Biology, 1272. pp.91-106. ISSN 1940-6029 (Book Chapter _OA Not required)
Full text not available from this repository.Abstract
Orphan G protein-coupled receptors represent an underexploited resource for drug discovery but pose a considerable challenge for assay development because their cognate G protein signaling pathways are often unknown. In this methodological chapter, we describe the use of constitutive activity, that is, the inherent ability of receptors to couple to their cognate G proteins in the absence of ligand, to inform the development of high-throughput screening assays for a particular orphan receptor. We specifically focus on a two-step process, whereby constitutive G protein coupling is first determined using yeast Gpa1/human G protein chimeras linked to growth and β-galactosidase generation. Coupling selectivity is then confirmed in mammalian cells expressing endogenous G proteins and driving accumulation of transcription factor-fused luciferase reporters specific to each of the classes of G protein. Based on these findings, high-throughput screening campaigns can be performed on the already miniaturized mammalian reporter system.
| Item Type: | Article |
|---|---|
| Subjects: | R Medicine > R Medicine (General) |
| Depositing User: | Repository Administrator |
| Date Deposited: | 05 Feb 2016 01:38 |
| Last Modified: | 11 Jan 2019 03:23 |
| URI: | https://eprints.victorchang.edu.au/id/eprint/336 |
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