TonB protein appears to transduce energy by shuttling between the cytoplasmic membrane and the outer membrane in Escherichia coli

Mol Microbiol. 1997 Apr;24(2):271-83. doi: 10.1046/j.1365-2958.1997.3331703.x.


The energy source for active transport of iron-siderophore complexes and vitamin B12 across the outer membrane in Gram-negative bacteria is the cytoplasmic membrane proton-motive force (pmf). TonB protein is required in this process to transduce cytoplasmic membrane energy to the outer membrane. In this study, Escherichia coli TonB was found to be distributed in sucrose density gradients approximately equally between the cytoplasmic membrane and the outer membrane fractions, while two proteins with which it is known to interact, ExbB and ExbD, as well as the NADH oxidase activity characteristic of the cytoplasmic membrane, were localized in the cytoplasmic membrane fraction. Neither the N-terminus of TonB nor the cytoplasmic membrane pmf, both of which are essential for TonB activity, were required for TonB to associate with the outer membrane. When the TonB C-terminus was absent, TonB was found associated with the cytoplasmic membrane, suggesting that the C-terminus was required for outer membrane association. When ExbB and ExbD, as well as their cross-talk-competent homologues ToIQ and ToIR, were absent, TonB was found associated with the outer membrane. TetA-TonB protein, which cannot interact with ExbB/D, was likewise found associated with the outer membrane. These results indicated that the role of ExbB/D in energy transduction is to bring TonB that has reached the outer membrane back to associate with the cytoplasmic membrane. Two possible explanations exist for the observations presented in this study. One possibility is that TonB transduces energy by shuttling between membranes, and, at some stages in the energy-transduction cycle, is associated with either the cytoplasmic membrane or the outer membrane, but not with both at the same time. This hypothesis, together with the alternative interpretation that TonB remains localized in the cytoplasmic membrane and changes its affinity for the outer and cytoplasmic membrane during energy transduction, are incorporated with previous observations into two new models, consistent with the novel aspects of this system, that describe a mechanism for TonB-dependent energy transduction.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antiporters / metabolism
  • Bacterial Outer Membrane Proteins / analysis
  • Bacterial Outer Membrane Proteins / isolation & purification
  • Bacterial Proteins / metabolism*
  • Bacterial Proteins / physiology
  • Cell Membrane / chemistry*
  • Cell Membrane / metabolism*
  • Escherichia coli / chemistry*
  • Escherichia coli / metabolism*
  • Escherichia coli Proteins*
  • Membrane Proteins / metabolism*
  • Membrane Proteins / physiology
  • Multienzyme Complexes / metabolism
  • NADH, NADPH Oxidoreductases / metabolism
  • Proton-Motive Force


  • Antiporters
  • Bacterial Outer Membrane Proteins
  • Bacterial Proteins
  • Escherichia coli Proteins
  • ExbB protein, E coli
  • Membrane Proteins
  • Multienzyme Complexes
  • tetA protein, Bacteria
  • tonB protein, Bacteria
  • tonB protein, E coli
  • tolQ protein, E coli
  • tolR protein, E coli
  • exbD protein, E coli
  • NADH oxidase
  • NADH, NADPH Oxidoreductases