Title

Bioluminescence Resonance Energy Transfer (BRET).

Description

BRET is a non-radiative energy transfer technique used to study molecular interactions within cells. Its principle is based on the use of a bioluminescent enzyme, luciferase (Renilla, NanoLuc, Firefly, etc.), which acts as an energy donor.

When luciferase catalyzes a chemical reaction, it emits light. If a fluorescent molecule (acceptor such as Yellow Fluorescent Protein) is close enough to the donor, the light energy emitted by luciferase can be transferred non-radiatively to the acceptor, causing fluorescence emission at a specific wavelength.

This phenomenon only occurs if the two molecules are in close proximity (less than 10 nanometers), which makes it possible to detect and measure molecular interactions, such as those between proteins in real time in a cellular environment.

Applications

BRET has been widely used since the 2000s in biomedical research to study interactions between proteins, monitor cellular processes in real time and develop second messenger detection tests.

BRET is used on the platform to analyze protein-protein interactions in cells such as interactions between G protein-coupled receptors (GPCRs), the dissociation of heterotrimeric G protein complexes or the recruitment of effectors at the receptor level such as GRKs (G-protein Regulating Kinases) or b-arrestins1/2.

In addition, BRET allows the generation of biosensitive sensors such as CAMYEL (cAMP sensor using YFP-Epac-RLuc) which is capable of detecting changes in cAMP concentration in cells.