Projects per year
Abstract
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.
Original language | English |
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Article number | e84512 |
Journal | PLoS ONE |
Volume | 8 |
Issue number | 12 |
DOIs | |
Publication status | Published - 23 Dec 2013 |
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Dive into the research topics of 'Mutational and topological analysis of the Escherichia coli BamA protein'. Together they form a unique fingerprint.Projects
- 4 Finished
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Selective Biochemical and Synthetic biology approaches for improved delivery of recombinant proteins to the extracellular Milieu
Henderson, I.
Biotechnology & Biological Sciences Research Council
1/12/11 → 31/05/15
Project: Research Councils
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Structural Basis of the Outer Membrane Protein Assembly System by NMR Spectroscopy
Overduin, M., Henderson, I. & Knowles, T.
Biotechnology & Biological Sciences Research Council
1/12/09 → 30/11/13
Project: Research Councils
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Understanding the Role of the Bam Complex in the Biogenesis of Outer Membrane Proteins
Henderson, I. & Overduin, M.
1/01/09 → 31/12/11
Project: Research Councils
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POTRA Domain Structure and Function by NMR Spectroscopy
Overduin, M. & Henderson, I.
Biotechnology & Biological Sciences Research Council
1/10/07 → 30/09/10
Project: Research Councils