This technology consists in a set of more than 25 BioSensor assays, each designed to follow one specific signaling pathway. This BRET-based BioSensor platform was implemented to interrogate the signaling complexity associated with GPCR activation and is the tool of choice for the identification of biased ligands.
Unimolecular BRET-based BioSensors that are based on the detection of intramolecular rearrangements of a sensor attached to both energy donor and acceptor that is promoted either by the binding of a second messenger or partner protein, a translocation event from the cytoplasm to the plasma membrane or a phosphorylation.
The example presents a BioSensor that detects the production of cAMP. Binding of the receptor to the BioSensor leads to a conformational rearrangement that changes the distance between the energy donor luciferase (Luc) and acceptor GFP thus yielding a change in BRET.
Bimolecular BRET-based BioSensors that are based on protein–protein interactions that are modified by a signaling event.
The example presents a BioSensor monitoring the activation of a G-protein by assessing the separation between the Gα and Gγ subunits that follows G-protein activation and results in a BRET change.