Förster Resonance Energy Transfer Measurements in Living Plant Cells

10.3791/62758 ◽  
2021 ◽  
Author(s):  
Sonja Michèle Schmidtpott ◽  
Thorsten Seidel
Keyword(s):  
Energy Transfer ◽  
Resonance Energy Transfer ◽  
Plant Cells ◽  
Resonance Energy ◽  
Living Plant

scholarly journals Quantification of Förster resonance energy transfer by monitoring sensitized emission in living plant cells

2013 ◽  
Vol 4 ◽  
Author(s):  
Sara M. Müller ◽  
Helena Galliardt ◽  
Jessica Schneider ◽  
B. George Barisas ◽  
Thorsten Seidel
Keyword(s):  
Energy Transfer ◽  
Resonance Energy Transfer ◽  
Plant Cells ◽  
Resonance Energy ◽  
Forster Resonance Energy Transfer ◽  
Förster Resonance Energy Transfer ◽  
Living Plant ◽  
Förster Resonance

scholarly journals Förster resonance energy transfer demonstrates a flavonoid metabolon in living plant cells that displays competitive interactions between enzymes

FEBS Letters ◽  
2011 ◽  
Vol 585 (14) ◽  
pp. 2193-2198 ◽  
Author(s):  
Kevin C. Crosby ◽  
Anna Pietraszewska-Bogiel ◽  
Theodorus W.J. Gadella ◽  
Brenda S.J. Winkel
Keyword(s):  
Energy Transfer ◽  
Resonance Energy Transfer ◽  
Plant Cells ◽  
Resonance Energy ◽  
Forster Resonance Energy Transfer ◽  
Competitive Interactions ◽  
Förster Resonance Energy Transfer ◽  
Living Plant ◽  
Förster Resonance

scholarly journals Combined Bimolecular Fluorescence Complementation and Förster Resonance Energy Transfer Reveals Ternary SNARE Complex Formation in Living Plant Cells

2010 ◽  
Vol 152 (3) ◽  
pp. 1135-1147 ◽  
Author(s):  
Mark Kwaaitaal ◽  
Nana F. Keinath ◽  
Simone Pajonk ◽  
Christoph Biskup ◽  
Ralph Panstruga
Keyword(s):  
Energy Transfer ◽  
Complex Formation ◽  
Resonance Energy Transfer ◽  
Plant Cells ◽  
Resonance Energy ◽  
Snare Complex ◽  
Bimolecular Fluorescence Complementation ◽  
Living Plant ◽  
Snare Complex Formation ◽  
Fluorescence Complementation

scholarly journals In vivomonitoring of plant small GTPase activation using a Förster resonance energy transfer biosensor

2018 ◽  
Author(s):  
Hann Ling Wong ◽  
Akira Akamatsu ◽  
Qiong Wang ◽  
Masayuki Higuchi ◽  
Tomonori Matsuda ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Resonance Energy Transfer ◽  
Plant Cells ◽  
Resonance Energy ◽  
Small Gtpases ◽  
Small Gtpase ◽  
Forster Resonance Energy Transfer ◽  
Förster Resonance Energy Transfer ◽  
Living Plant ◽  
Förster Resonance

ABSTRACTSmall GTPases act as molecular switches that regulate various plant responses such as disease resistance, pollen tube growth, root hair development, cell wall patterning and hormone responses. Thus, to monitor their activation status within plant cells is believed to be the key step in understanding their roles. We have established a plant version of a Förster resonance energy transfer (FRET) probe called Ras and interacting protein chimeric unit (Raichu) that can successfully monitor activation of the rice small GTPase OsRac1 during various defence responses in rice cells. Here, we describe the protocol for visualizing spatiotemporal activity of plant Rac/ROP GTPase in living plant cells, transfection of rice protoplasts withRaichu-OsRac1and acquisition of FRET images. Our protocol should be widely adaptable for monitoring activation for other plant small GTPases and for other FRET sensors in various plant cells.

scholarly journals Three-fluorophore FRET enables the analysis of ternary protein association in living plant cells

2019 ◽  
Author(s):  
Nina Glöckner ◽  
Sven zur Oven-Krockhaus ◽  
Leander Rohr ◽  
Frank Wackenhut ◽  
Moritz Burmeister ◽  
...  
Keyword(s):  
Ternary Complex ◽  
Resonance Energy Transfer ◽  
Plant Cells ◽  
Resonance Energy ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Living Plant ◽  
Protein Protein Interaction ◽  
Lifetime Imaging ◽  
Dependent Cell

AbstractProtein-protein interaction studies provide valuable insights into cellular signaling. The well-studied brassinosteroid (BR) signaling is initiated by the hormone-binding receptor Brassinosteroid Insensitive 1 (BRI1) and its co-receptor BRI1 Associated Kinase 1 (BAK1). BRI1 and BAK1 were shown to interact independently with the Receptor-Like Protein 44 (RLP44), which is implicated in BRI1/BAK1-dependent cell wall integrity perception. To demonstrate the proposed complex formation of BRI1, BAK1 and RLP44, we established three-fluorophore intensity-based spectral Förster resonance energy transfer (FRET) and FRET-fluorescence lifetime imaging microscopy (FLIM) for living plant cells. Our evidence indicates that RLP44, BRI1 and BAK1 form a ternary complex in a distinct plasma membrane nanodomain. In contrast, although the immune receptor Flagellin Sensing 2 (FLS2) also forms a heteromer with BAK1, the FLS2/BAK1 complexes are localized in other nanodomains. In general, our three-fluorophore FRET approaches provide a feasible basis for studying the interaction and sub-compartmentalization of proteins in great detail.

Quantum Dot Bioconjugates as Energy Donors in Fluorescence Resonance Energy Transfer Assays

2003 ◽  
Vol 773 ◽  
Author(s):  
Aaron R. Clapp ◽  
Igor L. Medintz ◽  
J. Matthew Mauro ◽  
Hedi Mattoussi
Keyword(s):  
Energy Transfer ◽  
Quantum Dot ◽  
Organic Dyes ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Genetically Engineered ◽  
Molar Ratio ◽  
Binding Assays ◽  
Specific Sequence ◽  
Donor Acceptor

AbstractLuminescent CdSe-ZnS core-shell quantum dot (QD) bioconjugates were used as energy donors in fluorescent resonance energy transfer (FRET) binding assays. The QDs were coated with saturating amounts of genetically engineered maltose binding protein (MBP) using a noncovalent immobilization process, and Cy3 organic dyes covalently attached at a specific sequence to MBP were used as energy acceptor molecules. Energy transfer efficiency was measured as a function of the MBP-Cy3/QD molar ratio for two different donor fluorescence emissions (different QD core sizes). Apparent donor-acceptor distances were determined from these FRET studies, and the measured distances are consistent with QD-protein conjugate dimensions previously determined from structural studies.

Sign in / Sign up

Export Citation Format

Share Document