title: Organization of the Escherichia coli aerobic enzyme complexes of oxidative phosphorylation in dynamic domains within the cytoplasmic membrane.
abstract: The Escherichia coli cytoplasmic membrane contains the enzyme complexes of oxidative phosphorylation (OXPHOS). Not much is known about their supramolecular organization and their dynamics within the membrane in this model organism. In mitochondria and other bacteria, it was demonstrated by nondenaturing electrophoretic methods and electron microscopy that the OXPHOS complexes are organized in so-called supercomplexes, stable assemblies with a defined number of the individual enzyme complexes. To investigate the organization of the E. coli enzyme complexes of aerobic OXPHOS in vivo, we established fluorescent protein fusions of the NADH:ubiquinone oxidoreductase, the succinate:ubiquinone oxidoreductase, the cytochrome bd-I, and the cytochrome bo3 terminal oxidases, and the FoF1 ATP-synthase. The fusions were integrated in the chromosome to prevent artifacts caused by protein overproduction. Biochemical analysis revealed that all modified complexes were fully assembled, active, and stable. The distribution of the OXPHOS complexes in living cells was determined using total internal reflection fluorescence microscopy. The dynamics within the membrane were detected by fluorescence recovery after photobleaching. All aerobic OXPHOS complexes showed an uneven distribution in large mobile patches within the E. coli cytoplasmic membrane. It is discussed whether the individual OXPHOS complexes are organized as clustered individual complexes, here called "segrazones."
authors.affiliation: Institut für Biochemie, Albert-Ludwigs-Universität, Albertstraße 21, Freiburg, 79104, Germany.
authors.forename: Heiko
authors.initials: H
authors.lastname: Erhardt
authors.forename: Felix
authors.initials: F
authors.lastname: Dempwolff
authors.forename: Moritz
authors.initials: M
authors.lastname: Pfreundschuh
authors.forename: Marc
authors.initials: M
authors.lastname: Riehle
authors.forename: Caspar
authors.initials: C
authors.lastname: Schäfer
authors.forename: Thomas
authors.initials: T
authors.lastname: Pohl
authors.forename: Peter
authors.initials: P
authors.lastname: Graumann
authors.forename: Thorsten
authors.initials: T
authors.lastname: Friedrich
mesh_terms: D000332:Aerobiosis; D019677:Artificial Gene Fusion; D002462:Cell Membrane / Q000201:enzymology*; D004926:Escherichia coli / Q000201:enzymology* / Q000378:metabolism*; D017930:Genes, Reporter; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D008856:Microscopy, Fluorescence; D009097:Multienzyme Complexes / Q000378:metabolism*; D010085:Oxidative Phosphorylation
chemical_list: D008164:Luminescent Proteins; D009097:Multienzyme Complexes
keywords: Escherichia coli; FRAP; TIRF microscopy; in vivo fluorescence microsocopy; membrane protein organization; oxidative phosphorylation
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