Subunit rotation in a single FoF1-ATP synthase in a living bacterium monitored by FRET

@inproceedings{Seyfert2011SubunitRI,
  title={Subunit rotation in a single FoF1-ATP synthase in a living bacterium monitored by FRET},
  author={Karin Seyfert and Takuya Oosaka and Hideyuki Yaginuma and Stefan Ernst and Hiroyuki Noji and Ryota Iino and Michael Boersch},
  booktitle={BiOS},
  year={2011}
}
FoF1-ATP synthase is the ubiquitous membrane-bound enzyme in mitochondria, chloroplasts and bacteria which provides the 'chemical energy currency' adenosine triphosphate (ATP) for cellular processes. In Escherichia coli ATP synthesis is driven by a proton motive force (PMF) comprising a proton concentration difference ΔpH plus an electric potential ΔΨ across the lipid membrane. Single-molecule in vitro experiments have confirmed that proton-driven subunit rotation within FoF1-ATP synthase is… 
Diffusion properties of single FoF1-ATP synthases in a living bacterium unraveled by localization microscopy
FoF1-ATP synthases in Escherichia coli (E. coli) bacteria are membrane-bound enzymes which use an internal protondriven rotary double motor to catalyze the synthesis of adenosine triphosphate (ATP).
Observing single FoF1-ATP synthase at work using an improved fluorescent protein mNeonGreen as FRET donor
TLDR
The novel FRET donor mNeonGreen is evaluated as a fusion to FoF1-ATP synthase and compare it to the previously used fluorophore EGFP to evaluate the biochemical purification procedures and activity measurements of the fully functional mutant enzyme.
Monitoring transient elastic energy storage within the rotary motors of single FoF1-ATP synthase by DCO-ALEX FRET
TLDR
The detection of reversible elastic deformations between the rotor parts of Fo and F1 is reported and the maximum angular displacement during the load-free rotation is estimated using Monte Carlo simulations.
Subunit rotation in single FRET-labeled F1-ATPase hold in solution by an anti-Brownian electrokinetic trap
TLDR
Monte Carlo simulations are used to reveal that stepwise FRET efficiency changes can be analyzed by Hidden Markov Models even at the limit of a low signal-to-background ratio that was expected due to high background count rates caused by the microfluidics of the ABELtrap.
Twisting and subunit rotation in single FOF1-ATP synthase
TLDR
Recent developments of approaches to monitor the step size of subunit rotation and the transient elastic energy storage mechanism in single FOF1-ATP synthases are reviewed.
Unraveling the Rotary Motors in FoF1-ATP Synthase by Time-Resolved Single-Molecule FRET
Detection of single fluorophore molecules was reported 25 years ago, at first in a crystalline matrix at cryogenic temperatures but quickly followed by single-molecule studies of biological machines
Microscopy of single FoF1‐ATP synthases— The unraveling of motors, gears, and controls
TLDR
Förster resonance energy transfer, which has been used for simultaneous monitoring of conformational changes of different parts of this rotary motor, is one of them and may become the tool for the analysis of single FoF1‐ATP synthases in membranes of living cells.
Observing conformations of single FoF1-ATP synthases in a fast anti-Brownian electrokinetic trap
TLDR
A version of an ABELtrap with a laser focus pattern generated by electro-optical beam deflectors and controlled by a programmable FPGA is presented, which could hold single fluorescent nanobeads for more than 100 seconds and increase the observation times of a single particle more than 1000-fold.
Binding of the immunomodulatory drug Bz-423 to mitochondrial FoF1-ATP synthase in living cells by FRET acceptor photobleaching
TLDR
This work measured uptake and binding of a Cy5-labeled Bz-423 derivative to mitochondrial FoF1-ATP synthase in living yeast cells using FRET acceptor photobleaching microscopy and confirmed the binding of Cy5 to the top of the F1 domain of the enzyme in mitochondria of living Saccharomyces cerevisiae cells.
3D-localization microscopy and tracking of FoF1-ATP synthases in living bacteria
TLDR
The one-dimensional diffusion coefficient of FoF1-ATP synthase diffusing on the long axis in living E. coli cells is obtained and the limited size of the observation area in the membrane with its significant membrane curvature has to be considered.
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