title: Energy-dependent conformational change in the TolA protein of Escherichia coli involves its N-terminal domain, TolQ, and TolR.
abstract: TolQ, TolR, and TolA inner membrane proteins of Escherichia coli are involved in maintaining the stability of the outer membrane. They share homology with the ExbB, ExbD, and TonB proteins, respectively. The last is involved in energy transduction between the inner and the outer membrane, and its conformation has been shown to depend on the presence of the proton motive force (PMF), ExbB, and ExbD. Using limited proteolysis experiments, we investigated whether the conformation of TolA was also affected by the PMF. We found that dissipation of the PMF by uncouplers led to the formation of a proteinase K digestion fragment of TolA not seen when uncouplers are omitted. This fragment was also detected in Delta tolQ, Delta tolR, and tolA(H22P) mutants but, in contrast to the parental strain, was also seen in the absence of uncouplers. We repeated those experiments in outer membrane mutants such as lpp, pal, and Delta rfa mutants: the behavior of TolA in lpp mutants was similar to that observed with the parental strain. However, the proteinase K-resistant fragment was never detected in the Delta rfa mutant. Altogether, these results suggest that TolA is able to undergo a PMF-dependent change of conformation. This change requires TolQ, TolR, and a functional TolA N-terminal domain. The potential role of this energy-dependent process in the stability of the outer membrane is discussed.
authors: Unité de Microbiologie et Génétique, ERS2009 (CNRS-INSA-Université Lyon 1), F-69622 Villeurbanne Cedex, France.
authors: P
authors: P
authors: Germon
authors: M C
authors: MC
authors: Ray
authors: A
authors: A
authors: Vianney
authors: J C
authors: JC
authors: Lazzaroni
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002352:Carrier Proteins / Q000235:genetics; D019286:Endopeptidase K / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism / Q000502:physiology; D029968:Escherichia coli Proteins; D008074:Lipoproteins / Q000235:genetics; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D016296:Mutagenesis; D010457:Peptidoglycan / Q000235:genetics; D033903:Periplasmic Proteins; D011487:Protein Conformation; D011509:Proteoglycans; D018892:Proton-Motive Force / Q000502:physiology*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; C031163:ExcC protein, E coli; D008074:Lipoproteins; C099932:Lpp protein, E coli; D008565:Membrane Proteins; D010457:Peptidoglycan; D033903:Periplasmic Proteins; D011509:Proteoglycans; C062093:tolA protein, E coli; C062094:tolB protein, E coli; C060792:tolQ protein, E coli; C060794:tolR protein, E coli; C069988:PplA protein, Legionella pneumophila; D019286:Endopeptidase K