A flow cytometric method to detect protein-protein interaction in living cells by directly visualizing donor fluorophore quenching during CFP?YFP fluorescence resonance energy transfer (FRET)

Cytometry ◽  
2003 ◽  
Vol 55A (2) ◽  
pp. 71-85 ◽  
Author(s):  
Liusheng He ◽  
Douglas P. Olson ◽  
Xiaoli Wu ◽  
Tatiana S. Karpova ◽  
James G. McNally ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Protein Interaction ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Living Cells ◽  
Protein Protein Interaction ◽  
Flow Cytometric Method ◽  
Flow Cytometric ◽  
Fluorescence Resonance

scholarly journals A Fluorescence Resonance Energy Transfer-Based Analytical Tool for Nitrate Quantification in Living Cells

ACS Omega ◽  
2020 ◽  
Vol 5 (46) ◽  
pp. 30306-30314
Author(s):  
Urooj Fatima ◽  
Fuad Ameen ◽  
Neha Soleja ◽  
Parvez Khan ◽  
Abobakr Almansob ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Living Cells ◽  
Analytical Tool ◽  
Fluorescence Resonance

scholarly journals Visualization of ATP levels inside single living cells with fluorescence resonance energy transfer-based genetically encoded indicators

2009 ◽  
Vol 106 (37) ◽  
pp. 15651-15656 ◽  
Author(s):  
Hiromi Imamura ◽  
Kim P. Huynh Nhat ◽  
Hiroko Togawa ◽  
Kenta Saito ◽  
Ryota Iino ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Atp Synthase ◽  
Hela Cells ◽  
Dissociation Constants ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Living Cells ◽  
Atp Levels ◽  
Fluorescence Resonance

Adenosine 5′-triphosphate (ATP) is the major energy currency of cells and is involved in many cellular processes. However, there is no method for real-time monitoring of ATP levels inside individual living cells. To visualize ATP levels, we generated a series of fluorescence resonance energy transfer (FRET)-based indicators for ATP that were composed of the ε subunit of the bacterial FoF1-ATP synthase sandwiched by the cyan- and yellow-fluorescent proteins. The indicators, named ATeams, had apparent dissociation constants for ATP ranging from 7.4 μM to 3.3 mM. By targeting ATeams to different subcellular compartments, we unexpectedly found that ATP levels in the mitochondrial matrix of HeLa cells are significantly lower than those of cytoplasm and nucleus. We also succeeded in measuring changes in the ATP level inside single HeLa cells after treatment with inhibitors of glycolysis and/or oxidative phosphorylation, revealing that glycolysis is the major ATP-generating pathway of the cells grown in glucose-rich medium. This was also confirmed by an experiment using oligomycin A, an inhibitor of FoF1-ATP synthase. In addition, it was demonstrated that HeLa cells change ATP-generating pathway in response to changes of nutrition in the environment.

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