2019
DOI: 10.3390/molecules24030537
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Bioluminescence Resonance Energy Transfer as a Method to Study Protein-Protein Interactions: Application to G Protein Coupled Receptor Biology

Abstract: The bioluminescence resonance energy transfer (BRET) approach involves resonance energy transfer between a light-emitting enzyme and fluorescent acceptors. The major advantage of this technique over biochemical methods is that protein-protein interactions (PPI) can be monitored without disrupting the natural environment, frequently altered by detergents and membrane preparations. Thus, it is considered as one of the most versatile technique for studying molecular interactions in living cells at “physiological”… Show more

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Cited by 41 publications
(37 citation statements)
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“…Besides allowing to avoid the disruption of the natural environment frequently altered in classical biochemical methods using detergents and membrane preparations, BRET is useful to discriminate whether the interactions occur in cis (in the same cell) or in trans (between two cells) orientation 27 . For this purpose, the formation of homo‐ or heterocomplexes of LINGO homologs was examined in living HEK‐293 cells using quantitative BRET analysis and the proper controls required by this approach 30,31 . After transfection of HEK‐293 cells with two LINGO constructs carrying in their C‐terminus end either the yellow fluorescent protein (LINGO‐1‐YFP; LINGO‐2‐YFP, LINGO‐3‐YFP, or LINGO‐4‐YFP) or the Renilla luciferase (LINGO‐1‐Rluc, LINGO‐2‐RLuc, LINGO‐3‐RLuc, or LINGO‐4 RLuc), we observed a specific BRET signal generated by the formation of LINGO dimer/oligomers in living cells (Figure 6).…”
Section: Resultsmentioning
confidence: 99%
“…Besides allowing to avoid the disruption of the natural environment frequently altered in classical biochemical methods using detergents and membrane preparations, BRET is useful to discriminate whether the interactions occur in cis (in the same cell) or in trans (between two cells) orientation 27 . For this purpose, the formation of homo‐ or heterocomplexes of LINGO homologs was examined in living HEK‐293 cells using quantitative BRET analysis and the proper controls required by this approach 30,31 . After transfection of HEK‐293 cells with two LINGO constructs carrying in their C‐terminus end either the yellow fluorescent protein (LINGO‐1‐YFP; LINGO‐2‐YFP, LINGO‐3‐YFP, or LINGO‐4‐YFP) or the Renilla luciferase (LINGO‐1‐Rluc, LINGO‐2‐RLuc, LINGO‐3‐RLuc, or LINGO‐4 RLuc), we observed a specific BRET signal generated by the formation of LINGO dimer/oligomers in living cells (Figure 6).…”
Section: Resultsmentioning
confidence: 99%
“…Furimazine has minimal background bioluminescence compared to other luciferins [ 28 ], thus, exhibiting a higher signal-to-background ratio. All these properties of NanoLuc have led to its utility in a variety of applications including resonance energy transfer-based protein–protein interaction determination (BRET), gene regulation, and monitoring protein stability in diseased conditions [ 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 ]. In fact, a number of biosensing strategies have been devised based on the BRET phenomenon [ 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 ].…”
Section: Introductionmentioning
confidence: 99%
“…Protein complexes are usually identified by characterization of PPI. Some of the commonly used experimental strategies to detect PPIs, include the yeast two-hybrid system (Y2H; Paiano et al, 2019 ), protein-fragment complementation assay (PCA; Tarassov et al, 2008 ; Michnick et al, 2010 ), fluorescence resonance energy transfer (FRET; El Khamlichi et al, 2019 ), and affinity purification plus mass spectrometry (AP-MS; Gavin et al, 2006 ; Krogan et al, 2006 ; Volkel et al, 2010 ; Babu et al, 2012 ; Adelmant et al, 2019 ). These methods help us collect a vast set of protein-protein interaction data from different organisms.…”
Section: Proteome-scale Analyses Of Protein Complexesmentioning
confidence: 99%