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Structural and functional dissection reveals distinct roles of Ca(2+)-binding sites in the giant adhesin SiiE of Salmonella enterica
The giant non-fimbrial adhesin SiiE of Salmonella enterica mediates the first contact to the apical site of epithelial cells and enables subsequent invasion. SiiE is a 595 kDa protein composed of 53 repetitive bacterial immunoglobulin (BIg) domains and the only known substrate of the SPI4-encoded ty...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5466336/ https://www.ncbi.nlm.nih.gov/pubmed/28558023 http://dx.doi.org/10.1371/journal.ppat.1006418 |
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author | Peters, Britta Stein, Johanna Klingl, Stefan Sander, Nathalie Sandmann, Achim Taccardi, Nicola Sticht, Heinrich Gerlach, Roman G. Muller, Yves A. Hensel, Michael |
author_facet | Peters, Britta Stein, Johanna Klingl, Stefan Sander, Nathalie Sandmann, Achim Taccardi, Nicola Sticht, Heinrich Gerlach, Roman G. Muller, Yves A. Hensel, Michael |
author_sort | Peters, Britta |
collection | PubMed |
description | The giant non-fimbrial adhesin SiiE of Salmonella enterica mediates the first contact to the apical site of epithelial cells and enables subsequent invasion. SiiE is a 595 kDa protein composed of 53 repetitive bacterial immunoglobulin (BIg) domains and the only known substrate of the SPI4-encoded type 1 secretion system (T1SS). The crystal structure of BIg50-52 of SiiE revealed two distinct Ca(2+)-binding sites per BIg domain formed by conserved aspartate or glutamate residues. In a mutational analysis Ca(2+)-binding sites were disrupted by aspartate to serine exchange at various positions in the BIg domains of SiiE. Amounts of secreted SiiE diminish with a decreasing number of intact Ca(2+)-binding sites. BIg domains of SiiE contain distinct Ca(2+)-binding sites, with type I sites being similar to other T1SS-secreted proteins and type II sites newly identified in SiiE. We functionally and structurally dissected the roles of type I and type II Ca(2+)-binding sites in SiiE, as well as the importance of Ca(2+)-binding sites in various positions of SiiE. Type I Ca(2+)-binding sites were critical for efficient secretion of SiiE and a decreasing number of type I sites correlated with reduced secretion. Type II sites were less important for secretion, stability and surface expression of SiiE, however integrity of type II sites in the C-terminal portion was required for the function of SiiE in mediating adhesion and invasion. |
format | Online Article Text |
id | pubmed-5466336 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-54663362017-06-22 Structural and functional dissection reveals distinct roles of Ca(2+)-binding sites in the giant adhesin SiiE of Salmonella enterica Peters, Britta Stein, Johanna Klingl, Stefan Sander, Nathalie Sandmann, Achim Taccardi, Nicola Sticht, Heinrich Gerlach, Roman G. Muller, Yves A. Hensel, Michael PLoS Pathog Research Article The giant non-fimbrial adhesin SiiE of Salmonella enterica mediates the first contact to the apical site of epithelial cells and enables subsequent invasion. SiiE is a 595 kDa protein composed of 53 repetitive bacterial immunoglobulin (BIg) domains and the only known substrate of the SPI4-encoded type 1 secretion system (T1SS). The crystal structure of BIg50-52 of SiiE revealed two distinct Ca(2+)-binding sites per BIg domain formed by conserved aspartate or glutamate residues. In a mutational analysis Ca(2+)-binding sites were disrupted by aspartate to serine exchange at various positions in the BIg domains of SiiE. Amounts of secreted SiiE diminish with a decreasing number of intact Ca(2+)-binding sites. BIg domains of SiiE contain distinct Ca(2+)-binding sites, with type I sites being similar to other T1SS-secreted proteins and type II sites newly identified in SiiE. We functionally and structurally dissected the roles of type I and type II Ca(2+)-binding sites in SiiE, as well as the importance of Ca(2+)-binding sites in various positions of SiiE. Type I Ca(2+)-binding sites were critical for efficient secretion of SiiE and a decreasing number of type I sites correlated with reduced secretion. Type II sites were less important for secretion, stability and surface expression of SiiE, however integrity of type II sites in the C-terminal portion was required for the function of SiiE in mediating adhesion and invasion. Public Library of Science 2017-05-30 /pmc/articles/PMC5466336/ /pubmed/28558023 http://dx.doi.org/10.1371/journal.ppat.1006418 Text en © 2017 Peters et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Peters, Britta Stein, Johanna Klingl, Stefan Sander, Nathalie Sandmann, Achim Taccardi, Nicola Sticht, Heinrich Gerlach, Roman G. Muller, Yves A. Hensel, Michael Structural and functional dissection reveals distinct roles of Ca(2+)-binding sites in the giant adhesin SiiE of Salmonella enterica |
title | Structural and functional dissection reveals distinct roles of Ca(2+)-binding sites in the giant adhesin SiiE of Salmonella enterica |
title_full | Structural and functional dissection reveals distinct roles of Ca(2+)-binding sites in the giant adhesin SiiE of Salmonella enterica |
title_fullStr | Structural and functional dissection reveals distinct roles of Ca(2+)-binding sites in the giant adhesin SiiE of Salmonella enterica |
title_full_unstemmed | Structural and functional dissection reveals distinct roles of Ca(2+)-binding sites in the giant adhesin SiiE of Salmonella enterica |
title_short | Structural and functional dissection reveals distinct roles of Ca(2+)-binding sites in the giant adhesin SiiE of Salmonella enterica |
title_sort | structural and functional dissection reveals distinct roles of ca(2+)-binding sites in the giant adhesin siie of salmonella enterica |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5466336/ https://www.ncbi.nlm.nih.gov/pubmed/28558023 http://dx.doi.org/10.1371/journal.ppat.1006418 |
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