Single-molecule in vivo imaging of bacterial respiratory complexes indicates delocalized oxidative phosphorylation

Isabel Llorente-Garcia, Tchern Lenn, Heiko Erhardt, Oliver L. Harriman, Lu Ning Liu, Alex Robson, Sheng Wen Chiu, Sarah Matthews, Nicky J. Willis, Christopher D. Bray, Sang Hyuk Lee, Jae Yen Shin, Carlos Bustamante, Jan Liphardt, Thorsten Friedrich, Conrad W. Mullineaux, Mark C. Leake

Research output: Contribution to journalArticlepeer-review

80 Scopus citations


Chemiosmotic energy coupling through oxidative phosphorylation (OXPHOS) is crucial to life, requiring coordinated enzymes whose membrane organization and dynamics are poorly understood. We quantitatively explore localization, stoichiometry, and dynamics of key OXPHOS complexes, functionally fluorescent protein-tagged, in Escherichia coli using low-angle fluorescence and superresolution microscopy, applying single-molecule analysis and novel nanoscale co-localization measurements. Mobile 100-200 nm membrane domains containing tens to hundreds of complexes are indicated. Central to our results is that domains of different functional OXPHOS complexes do not co-localize, but ubiquinone diffusion in the membrane is rapid and long-range, consistent with a mobile carrier shuttling electrons between islands of different complexes. Our results categorically demonstrate that electron transport and proton circuitry in this model bacterium are spatially delocalized over the cell membrane, in stark contrast to mitochondrial bioenergetic supercomplexes. Different organisms use radically different strategies for OXPHOS membrane organization, likely depending on the stability of their environment.

Original languageEnglish (US)
Pages (from-to)811-824
Number of pages14
JournalBiochimica et Biophysica Acta - Bioenergetics
Issue number6
StatePublished - Jun 2014

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Cell Biology


  • Co-localization analysis
  • Cytoplasmic membrane
  • Fluorescence microscopy
  • Fluorescent protein
  • Oxidative phosphorylation
  • Single-molecule biophysics


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