Time-Resolved FRET Strategy to Screen GPCR Ligand Library
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Screening chemical libraries to find specific drugs for G protein-coupled receptors is still of major interest. Indeed, because of their major roles in all physiological functions, G protein-coupled receptors remain major targets for drug development programs. Currently, interest in GPCRs as drug targets has been boosted by the discovery of biased ligands, thus allowing the development of drugs not only specific for one target but also for the specific signaling cascade expected to have the therapeutic effect. Such molecules are then expected to display fewer side effects. To reach such a goal, there is much interest in novel, efficient, simple, and direct screening assays that may help identify any drugs interacting with the target, these being then analyzed for their biased activity. Here, we present an efficient strategy to screen ligands on their binding properties. The method described is based on time-resolved FRET between a receptor and a ligand. This method has already been used to develop new assays called Tag-lite® binding assays for numerous G protein-coupled receptors, proving its broad application and its power.
Key wordsTag-lite® screening G protein-coupled receptor Fluorescent ligand Time-resolved FRET Lanthanide Terbium Self-labeling enzyme Binding experiment
The development of the technique has been supported by the Fonds Unique Interministériel and OSEO in a collaborative program named “CELL2Lead.” Thanks are due to the Plateforme ARPÈGE of the Institut de Génomique Fonctionnelle.
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