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GloSensor assay for discovery of GPCR-selective ligands
Boda Arun Kumar1, Poonam Kumari1, Chandan Sona2
1Division of Pharmacology, CSIR-Central Drug Research Institute, Lucknow, India.
Methods in Cell Biology
|October 2, 2017
Summary
This chapter details the GloSensor assay for measuring cyclic adenosine monophosphate (cAMP) levels. It provides a cost-effective method for screening drug candidates targeting G protein-coupled receptors (GPCRs).
Area of Science:
- Biochemistry
- Pharmacology
- Molecular Biology
Background:
- G protein-coupled receptors (GPCRs) are crucial physiological modulators and prime therapeutic targets for diverse diseases.
- High-throughput functional assays are essential for cost-effective screening of large compound libraries to identify GPCR modulators.
- Existing fluorescence or chemiluminescence-based assays aid in GPCR research in academic and industrial settings.
Purpose of the Study:
- To provide a detailed, step-by-step protocol for performing cyclic adenosine monophosphate (cAMP) measurements using the GloSensor assay.
- To offer guidelines for the analysis and interpretation of results generated by the GloSensor assay.
- To illustrate the application of the GloSensor assay with examples of Gs- and Gi-coupled GPCRs.
Main Methods:
- Utilizes the GloSensor cAMP assay, a luminescence-based functional assay.
- Focuses on the practical implementation and execution of the assay protocol.
- Includes methods for data analysis and result interpretation.
Main Results:
- The chapter provides a comprehensive guide to conducting cAMP measurements using the GloSensor assay.
- Demonstrates the assay's utility in characterizing GPCR activity, including Gs- and Gi-coupled receptors.
- Offers practical insights into analyzing and interpreting assay data for drug discovery.
Conclusions:
- The GloSensor assay offers a versatile and effective method for functional screening of GPCRs.
- This assay facilitates the identification of agonists, antagonists, and allosteric modulators in a cost-effective manner.
- The provided guidelines and examples enhance the application of cAMP measurement in GPCR research and drug development.
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