Probing intermolecular protein-protein interactions in the calcium-sensing receptor homodimer using bioluminescence resonance energy transfer (BRET)

2002 ◽  
Vol 269 (20) ◽  
pp. 5076-5087 ◽  
Author(s):  
Anders A. Jensen ◽  
Jakob L. Hansen ◽  
Søren P. Sheikh ◽  
Hans Bräuner-Osborne
Keyword(s):  
Energy Transfer ◽  
Protein Interactions ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Bioluminescence Resonance Energy Transfer ◽  
Protein Protein Interactions ◽  
Calcium Sensing Receptor ◽  
Calcium Sensing

Bioluminescence resonance energy transfer–based imaging of protein–protein interactions in living cells

2019 ◽  
Vol 14 (4) ◽  
pp. 1084-1107 ◽  
Author(s):  
Hiroyuki Kobayashi ◽  
Louis-Philippe Picard ◽  
Anne-Marie Schönegge ◽  
Michel Bouvier
Keyword(s):  
Energy Transfer ◽  
Protein Interactions ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Living Cells ◽  
Bioluminescence Resonance Energy Transfer ◽  
Protein Protein Interactions

scholarly journals Bioluminescence Resonance Energy Transfer System for Measuring Dynamic Protein-Protein Interactions in Bacteria

mBio ◽  
2014 ◽  
Vol 5 (3) ◽  
Author(s):  
Boyu Cui ◽  
Yao Wang ◽  
Yunhong Song ◽  
Tietao Wang ◽  
Changfu Li ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Real Time ◽  
Protein Interactions ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Low Ph ◽  
Bacterial Cells ◽  
Bioluminescence Resonance Energy Transfer ◽  
Protein Protein Interactions ◽  
Excitation Light

ABSTRACT Protein-protein interactions are important for virtually every biological process, and a number of elegant approaches have been designed to detect and evaluate such interactions. However, few of these methods allow the detection of dynamic and real-time protein-protein interactions in bacteria. Here we describe a bioluminescence resonance energy transfer (BRET) system based on the bacterial luciferase LuxAB. We found that enhanced yellow fluorescent protein (eYFP) accepts the emission from LuxAB and emits yellow fluorescence. Importantly, BRET occurred when LuxAB and eYFP were fused, respectively, to the interacting protein pair FlgM and FliA. Furthermore, we observed sirolimus (i.e., rapamycin)-inducible interactions between FRB and FKBP12 and a dose-dependent abolishment of such interactions by FK506, the ligand of FKBP12. Using this system, we showed that osmotic stress or low pH efficiently induced multimerization of the regulatory protein OmpR and that the multimerization induced by low pH can be reversed by a neutralizing agent, further indicating the usefulness of this system in the measurement of dynamic interactions. This method can be adapted to analyze dynamic protein-protein interactions and the importance of such interactions in bacterial processes such as development and pathogenicity. IMPORTANCE Real-time measurement of protein-protein interactions in prokaryotes is highly desirable for determining the roles of protein complex in the development or virulence of bacteria, but methods that allow such measurement are not available. Here we describe the development of a bioluminescence resonance energy transfer (BRET) technology that meets this need. The use of endogenous excitation light in this strategy circumvents the requirement for the sophisticated instrument demanded by standard fluorescence resonance energy transfer (FRET). Furthermore, because the LuxAB substrate decanal is membrane permeable, the assay can be performed without lysing the bacterial cells, thus allowing the detection of protein-protein interactions in live bacterial cells. This BRET system added another useful tool to address important questions in microbiological studies.

Extended bioluminescence resonance energy transfer (eBRET) for monitoring prolonged protein–protein interactions in live cells

2006 ◽  
Vol 18 (10) ◽  
pp. 1664-1670 ◽  
Author(s):  
Kevin D.G. Pfleger ◽  
Jasmin R. Dromey ◽  
Matthew B. Dalrymple ◽  
Esther M.L. Lim ◽  
Walter G. Thomas ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Protein Interactions ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Live Cells ◽  
Bioluminescence Resonance Energy Transfer ◽  
Protein Protein Interactions
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