Confined Mobility of TonB and FepA in Escherichia coli Membranes.

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: eCollection Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science

Gene Expression Regulation, Bacterial*; Bacterial Outer Membrane Proteins/*metabolism ; Carrier Proteins/*metabolism ; Cell Membrane/*metabolism ; Escherichia coli/*metabolism ; Escherichia coli Proteins/*metabolism ; Membrane Proteins/*metabolism ; Receptors, Cell Surface/*metabolism; Antibodies, Neutralizing/pharmacology ; Bacterial Outer Membrane Proteins/antagonists & inhibitors ; Bacterial Outer Membrane Proteins/genetics ; Carrier Proteins/antagonists & inhibitors ; Carrier Proteins/genetics ; Cell Membrane/ultrastructure ; Cytoskeleton/metabolism ; Cytoskeleton/ultrastructure ; Diffusion ; Escherichia coli/genetics ; Escherichia coli/ultrastructure ; Escherichia coli Proteins/genetics ; Gene Deletion ; Membrane Proteins/genetics ; Molecular Dynamics Simulation ; Protein Binding ; Protein Transport ; Receptors, Cell Surface/antagonists & inhibitors ; Receptors, Cell Surface/genetics ; Single Molecule Imaging ; Time-Lapse Imaging

The important process of nutrient uptake in Escherichia coli, in many cases, involves transit of the nutrient through a class of beta-barrel proteins in the outer membrane known as TonB-dependent transporters (TBDTs) and requires interaction with the inner membrane protein TonB. Here we have imaged the mobility of the ferric enterobactin transporter FepA and TonB by tracking them in the membranes of live E. coli with single-molecule resolution at time-scales ranging from milliseconds to seconds. We employed simple simulations to model/analyze the lateral diffusion in the membranes of E.coli, to take into account both the highly curved geometry of the cell and artifactual effects expected due to finite exposure time imaging. We find that both molecules perform confined lateral diffusion in their respective membranes in the absence of ligand with FepA confined to a region [Formula: see text] μm in radius in the outer membrane and TonB confined to a region [Formula: see text] μm in radius in the inner membrane. The diffusion coefficient of these molecules on millisecond time-scales was estimated to be [Formula: see text] μm2/s and [Formula: see text] μm2/s for FepA and TonB, respectively, implying that each molecule is free to diffuse within its domain. Disruption of the inner membrane potential, deletion of ExbB/D from the inner membrane, presence of ligand or antibody to FepA and disruption of the MreB cytoskeleton was all found to further restrict the mobility of both molecules. Results are analyzed in terms of changes in confinement size and interactions between the two proteins.
Competing Interests: The authors have declared that no competing interests exist.

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R01 GM053836 United States GM NIGMS NIH HHS; R21 AI115187 United States AI NIAID NIH HHS

0 (Antibodies, Neutralizing)
0 (Bacterial Outer Membrane Proteins)
0 (Carrier Proteins)
0 (Escherichia coli Proteins)
0 (ExbB protein, E coli)
0 (Membrane Proteins)
0 (Receptors, Cell Surface)
0 (enterobactin receptor)
0 (tonB protein, E coli)
123424-75-9 (exbD protein, E coli)
149255-61-8 (MreB protein, E coli)

Date Created: 20161210 Date Completed: 20170705 Latest Revision: 20201215

20240628

PMC5147803

10.1371/journal.pone.0160862

27935943