Bioluminescence resonance energy transfer to monitor protein-protein interactions.
The bioluminescence resonance energy transfer (BRET) methodology allows for the study of protein-protein interactions as well as conformational changes within proteins or molecular complexes. BRET is a highly versatile technique that can be applied to in vitro studies using purified proteins, crude cell membranes, cell fractions obtained by centrifugation on a density gradient, as well as permeabilized cells. Importantly, BRET also allows for monitoring of protein-protein interactions, in real time, in intact living cells that can be submitted to various stimuli. Moreover, quantitative BRET analysis also permits a pharmacological approach of protein-protein interactions, allowing one to determine whether a given stimulus induces a conformational change within preassociated partners or increases the association (recruitment) between two separated partners. Determination of the proportion of the dimeric vs monomeric form of a protein in the cell also is possible. Therefore, the BRET technology can be considered as a new and powerful tool in the field of protein-protein interactions.
Issad
Tarik
T
Institut Cochin, INSERM, CNRS, Université Paris, France.
Jockers
Ralf
R
D000818:Animals; D002458:Cell Fractionation; D002460:Cell Line; D018076:DNA, Complementary / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006801:Humans; D049449:Luminescence; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*
D018076:DNA, Complementary; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins