Super resolution fluorescence microscopy and tracking of bacterial Flotillin (Reggie) paralogs provide evidence for defined-sized protein microdomains within the bacterial membrane but absence of clusters containing detergent-resistant proteins

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2016
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Dempwolff, Felix
Schmidt, Felix K.
Hervás, Ana B.
Stroh, Alex
Rösch, Thomas C.
Riese, Cornelius N.
Dersch, Simon
Heimerl, Thomas
Lucena, Daniella
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PLOS Genetics. 2016, 12(6), e1006116. ISSN 1553-7390. eISSN 1553-7404. Available under: doi: 10.1371/journal.pgen.1006116
Zusammenfassung

Biological membranes have been proposed to contain microdomains of a specific lipid composition, in which distinct groups of proteins are clustered. Flotillin-like proteins are conserved between pro-and eukaryotes, play an important function in several eukaryotic and bacterial cells, and define in vertebrates a type of so-called detergent-resistant microdomains. Using STED microscopy, we show that two bacterial flotillins, FloA and FloT, form defined assemblies with an average diameter of 85 to 110 nm in the model bacterium Bacillus subtilis. Interestingly, flotillin microdomains are of similar size in eukaryotic cells. The soluble domains of FloA form higher order oligomers of up to several hundred kDa in vitro, showing that like eukaryotic flotillins, bacterial assemblies are based in part on their ability to self-oligomerize. However, B. subtilis paralogs show significantly different diffusion rates, and consequently do not colocalize into a common microdomain. Dual colour time lapse experiments of flotillins together with other detergent-resistant proteins in bacteria show that proteins colocalize for no longer than a few hundred milliseconds, and do not move together. Our data reveal that the bacterial membrane contains defined-sized protein domains rather than functional microdomains dependent on flotillins. Based on their distinct dynamics, FloA and FloT confer spatially distinguishable activities, but do not serve as molecular scaffolds.

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ISO 690DEMPWOLFF, Felix, Felix K. SCHMIDT, Ana B. HERVÁS, Alex STROH, Thomas C. RÖSCH, Cornelius N. RIESE, Simon DERSCH, Thomas HEIMERL, Daniella LUCENA, Nikola HÜLSBUSCH, Claudia STÜRMER, Norio TAKESHITA, Reinhard FISCHER, Bruno ECKHARDT, Peter L. GRAUMANN, 2016. Super resolution fluorescence microscopy and tracking of bacterial Flotillin (Reggie) paralogs provide evidence for defined-sized protein microdomains within the bacterial membrane but absence of clusters containing detergent-resistant proteins. In: PLOS Genetics. 2016, 12(6), e1006116. ISSN 1553-7390. eISSN 1553-7404. Available under: doi: 10.1371/journal.pgen.1006116
BibTex
@article{Dempwolff2016Super-34697,
  year={2016},
  doi={10.1371/journal.pgen.1006116},
  title={Super resolution fluorescence microscopy and tracking of bacterial Flotillin (Reggie) paralogs provide evidence for defined-sized protein microdomains within the bacterial membrane but absence of clusters containing detergent-resistant proteins},
  number={6},
  volume={12},
  issn={1553-7390},
  journal={PLOS Genetics},
  author={Dempwolff, Felix and Schmidt, Felix K. and Hervás, Ana B. and Stroh, Alex and Rösch, Thomas C. and Riese, Cornelius N. and Dersch, Simon and Heimerl, Thomas and Lucena, Daniella and Hülsbusch, Nikola and Stürmer, Claudia and Takeshita, Norio and Fischer, Reinhard and Eckhardt, Bruno and Graumann, Peter L.},
  note={Article Number: e1006116}
}
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