Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/97117
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Type: | Journal article |
Title: | Structural and biochemical analysis of a single amino-acid mutant of WzzBSF that alters lipopolysaccharide O-antigen chain length in shigella flexneri |
Author: | Chang, C. Tran, E. Ericsson, D. Casey, L. Lonhienne, T. Benning, F. Morona, R. Kobe, B. |
Citation: | PLoS One, 2015; 10(9):e0138266-1-e0138266-21 |
Publisher: | Public Library of Science |
Issue Date: | 2015 |
ISSN: | 1932-6203 1932-6203 |
Editor: | Dobson, R. |
Statement of Responsibility: | Chiung-Wen Chang, Elizabeth N. H. Tran, Daniel J. Ericsson, Lachlan W. Casey, Thierry Lonhienne, Friederike Benning, Renato Morona, Bostjan Kobe |
Abstract: | Lipopolysaccharide (LPS), a surface polymer of Gram-negative bacteria, helps bacteria survive in different environments and acts as a virulence determinant of host infection. The O-antigen (Oag) component of LPS exhibits a modal chain-length distribution that is controlled by polysaccharide co-polymerases (PCPs). The molecular basis of the regulation of Oag chain-lengths remains unclear, despite extensive mutagenesis and structural studies of PCPs from Escherichia coli and Shigella. Here, we identified a single mutation (A107P) of the Shigella flexneri WzzBSF, by a random mutagenesis approach, that causes a shortened Oag chain-length distribution in bacteria. We determined the crystal structures of the periplasmic domains of wild-type WzzBSF and the A107P mutant. Both structures form a highly similar open trimeric assembly in the crystals, and show a similar tendency to self-associate in solution. Binding studies by bio-layer interferometry reveal cooperative binding of very short (VS)-core-plus-O-antigen polysaccharide (COPS) to the periplasmic domains of both proteins, but with decreased affinity for the A107P mutant. Our studies reveal that subtle and localized structural differences in PCPs can have dramatic effects on LPS chain-length distribution in bacteria, for example by altering the affinity for the substrate, which supports the role of the structure of the growing Oag polymer in this process. |
Keywords: | Escherichia coli Shigella flexneri Lipopolysaccharides O Antigens Amino Acids Bacterial Proteins Escherichia coli Proteins Crystallography, X-Ray Gene Expression Regulation, Bacterial Mutagenesis Protein Structure, Tertiary Mutation |
Rights: | © 2015 Chang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited |
DOI: | 10.1371/journal.pone.0138266 |
Published version: | http://dx.doi.org/10.1371/journal.pone.0138266 |
Appears in Collections: | Aurora harvest 7 Molecular and Biomedical Science publications |
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File | Description | Size | Format | |
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hdl_97117.pdf | Published version | 2.91 MB | Adobe PDF | View/Open |
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