3D-localization microscopy and tracking of FoF1-ATP synthases in living bacteria

  title={3D-localization microscopy and tracking of FoF1-ATP synthases in living bacteria},
  author={Anja Renz and Marc Renz and Diana Kl{\"u}tsch and Gabriele Deckers-Hebestreit and Michael B{\"o}rsch},
  booktitle={Photonics West - Biomedical Optics},
FoF1-ATP synthases are membrane-embedded protein machines that catalyze the synthesis of adenosine triphosphate. Using photoactivation-based localization microscopy (PALM) in TIR-illumination as well as structured illumination microscopy (SIM), we explore the spatial distribution and track single FoF1-ATP synthases in living E. coli cells under physiological conditions at different temperatures. For quantitative diffusion analysis by mean-squared-displacement measurements, the limited size of… 
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Imaging cytochrome C oxidase and FoF1-ATP synthase in mitochondrial cristae of living human cells by FLIM and superresolution microscopy
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Refined method to study the posttranslational regulation of alternative oxidases from Arabidopsis thaliana in vitro.
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Low membrane fluidity triggers lipid phase separation and protein segregation in living bacteria
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Observing monomer: dimer transitions of neurotensin receptors 1 in single SMALPs by homoFRET and in an ABELtrap
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Ligand-induced oligomerization of the human GPCR neurotensin receptor 1 monitored in living HEK293T cells
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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).
Microscopy of single FoF1‐ATP synthases— The unraveling of motors, gears, and controls
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.
Imaging and quantification of trans-membrane protein diffusion in living bacteria.
A novel approach, IPODD (inverse projection of displacement distributions), is introduced to obtain diffusion coefficients from the usually obtained 2-D projected diffusion trajectories of the highly 3-D curved bacterial membrane to provide correct diffusion coefficients and allow unravelling of heterogeneously diffusing populations.
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A method to extract multiple diffusion coefficients from trajectories obtained from SMF data, using cumulative probability distributions (CPDs), and demonstrates the power of this approach by quantifying the heterogeneous diffusion of the bacterial membrane protein TatA, which forms a pore for the translocation of folded proteins.
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Subunit rotation in a single FoF1-ATP synthase in a living bacterium monitored by FRET
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Monitoring the rotary motors of single FoF1-ATP synthase by synchronized multi channel TCSPC
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Rotation of the ε-subunit during ATP hydrolysis was divided into three major steps and the stopping positions of ε resulted in three distinct FRET efficiency levels and FRET donor lifetimes and the position of the FRET donors at the asubunit was calculated.