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Published on: December 2, 2022
G protein-coupled receptor internalization assays in the high-content screening format
Dorothea Haasen1, Andreas Schnapp, Martin J Valler
1Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany.
Methods in Enzymology
|November 18, 2006
Summary
High-content screening (HCS) measures G protein-coupled receptor (GPCR) internalization using advanced imaging. This method aids in discovering drugs that target GPCRs, crucial for pharmaceutical development.
Area of Science:
- Pharmacology
- Cell Biology
- Drug Discovery
Background:
- G protein-coupled receptors (GPCRs) are key therapeutic targets in the pharmaceutical industry.
- Developing selective compounds to modulate GPCR function is a major focus.
- High-content screening (HCS) integrates microscopy and image analysis for cellular studies.
Purpose of the Study:
- To describe the measurement of GPCR internalization using HCS.
- To present robust assay formats for studying GPCR-ligand interactions.
- To evaluate GPCR internalization assays as a drug discovery tool.
Main Methods:
- Utilizing a high-throughput, confocal cellular imaging device for HCS.
- Measuring ligand-induced internalization of GPCRs.
- Employing GPCRs labeled with fluorophore-conjugated ligands or antibodies.
Main Results:
- Developed robust assay formats for GPCR internalization measurement.
- Demonstrated the utility of HCS for analyzing GPCR ligand activity.
- Internalization assays provide complementary pharmacological data.
Conclusions:
- GPCR internalization assays are valuable for medium/high-throughput screening.
- These assays facilitate the cellular activity determination of GPCR ligands.
- HCS offers a powerful approach for GPCR-focused drug discovery.
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01:21G-protein Coupled Receptors
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01:15G Protein-coupled Receptors
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