how can we detect the presence of a g coupled protein receptor getting activated when there is a signal molecule received. how can we prove it experimentally apart from measuring the level of secondary messengers?
Hello Nisha Reghu
Several methods are available for studying GPCR modulation namely, ligand-binding-based assays, measurements of guanine nucleotide exchange, and functional activity-based assays of receptors. Functional activity assays use either reporter-based assays or methods that monitor the intracellular concentrations of the second messengers like cAMP, inositol trisphosphate (IP3), and intracellular Ca2+ mobilization.
Reporter-based assays offer the advantage of wide linearity and sensitivity. Reporter gene system is a technology where a reporter gene is synthesized in response to activation of a specific signaling cascade of interests, followed by monitoring of the reporter protein expression by its enzymatic activities linked with a variety of colorimetric, fluorescent or luminescent read-outs.
You may want to refer to the article I have attached below for your reference. It may be helpful.
Article Luciferase Reporter Assay System for Deciphering GPCR Pathways
Best.
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