Observing single FoF1-ATP synthase at work using an improved fluorescent protein mNeonGreen as FRET donor

@inproceedings{Heitkamp2016ObservingSF,
  title={Observing single FoF1-ATP synthase at work using an improved fluorescent protein mNeonGreen as FRET donor},
  author={Thomas Heitkamp and Gabriele Deckers-Hebestreit and Michael B{\"o}rsch},
  booktitle={SPIE BiOS},
  year={2016}
}
Adenosine triphosphate (ATP) is the universal chemical energy currency for cellular activities provided mainly by the membrane enzyme FoF1-ATP synthase in bacteria, chloroplasts and mitochondria. Synthesis of ATP is accompanied by subunit rotation within the enzyme. Over the past 15 years we have developed a variety of single-molecule FRET (smFRET) experiments to monitor catalytic action of individual bacterial enzymes in vitro. By specifically labeling rotating and static subunits within a… 
Fast ATP-Dependent Subunit Rotation in Reconstituted FoF1-ATP Synthase Trapped in Solution.
TLDR
It is reported that kinetic monitoring of functional rotation can be prolonged from milliseconds to seconds by utilizing an anti-Brownian electrokinetic trap (ABEL trap), and broad distributions of ATP-dependent catalytic rates were revealed.
Fast ATP-dependent Subunit Rotation in Reconstituted FoF1-ATP Synthase Trapped in Solution
TLDR
It is reported that kinetic monitoring of functional rotation can be prolonged from milliseconds to seconds by utilizing an Anti-Brownian electrokinetic trap (ABEL trap), and broad distributions of ATP-dependent catalytic rates were revealed.
Imaging cytochrome C oxidase and FoF1-ATP synthase in mitochondrial cristae of living human cells by FLIM and superresolution microscopy
TLDR
Their mitochondrial nano-environment was investigated by FLIM and superresolution microscopy in living human cells and different lifetimes and anisotropy values were found.
Towards monitoring conformational changes of the GPCR neurotensin receptor 1 by single-molecule FRET
TLDR
G protein-coupled receptor 1 was found to be monomeric in liposomes, with a small fraction being dimeric and oligomeric, showing homoFRET, and agonist binding to NTSR1 was demonstrated by time-resolved single-molecule Förster resonance energy transfer (smFRET), using neurotensin labeled with the fluorophore ATTO594.
Single-Molecular Förster Resonance Energy Transfer Measurement on Structures and Interactions of Biomolecules
TLDR
Some representative examples of smFRET research of recent years are summarized to exhibit the versatility and note typical strategies to further improve the performance ofsmFRET measurement on different biomolecules.
Confining Brownian motion of single nanoparticles in an ABELtrap
TLDR
The ABELtrap is an active feedback system cancelling the nano-object’s Brownian motion by applying an electric field and it is shown how the induced electrokinetic forces confine the motion of nanoparticles and proteoliposomes to the centre of the trap.

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