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Mutational and topological analysis of the Escherichia coli BamA protein.

Browning, Douglas F and Matthews, Sophie A and Rossiter, Amanda E and Sevastsyanovich, Yanina R and Jeeves, Mark and Mason, Jessica L and Wells, Timothy J and Wardius, Catherine A and Knowles, Timothy J and Cunningham, Adam F and Bavro, Vassiliy N and Overduin, Michael and Henderson, Ian R (2013) 'Mutational and topological analysis of the Escherichia coli BamA protein.' PLoS One, 8 (12). e84512-e84512. ISSN 1932-6203

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The multi-protein β-barrel assembly machine (BAM) of Escherichia coli is responsible for the folding and insertion of β-barrel containing integral outer membrane proteins (OMPs) into the bacterial outer membrane. An essential component of this complex is the BamA protein, which binds unfolded β-barrel precursors via the five polypeptide transport-associated (POTRA) domains in its N-terminus. The C-terminus of BamA contains a β-barrel domain, which tethers BamA to the outer membrane and is also thought to be involved in OMP insertion. Here we mutagenize BamA using linker scanning mutagenesis and demonstrate that all five POTRA domains are essential for BamA protein function in our experimental system. Furthermore, we generate a homology based model of the BamA β-barrel and test our model using insertion mutagenesis, deletion analysis and immunofluorescence to identify β-strands, periplasmic turns and extracellular loops. We show that the surface-exposed loops of the BamA β-barrel are essential.

Item Type: Article
Uncontrolled Keywords: Escherichia coli; Bacterial Outer Membrane Proteins; Escherichia coli Proteins; Fluorescent Antibody Technique; Blotting, Western; Mutagenesis; Protein Structure, Tertiary; Plasmids; Models, Molecular
Divisions: Faculty of Science and Health
Faculty of Science and Health > Life Sciences, School of
SWORD Depositor: Elements
Depositing User: Elements
Date Deposited: 28 Jun 2021 10:37
Last Modified: 06 Jan 2022 15:00

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