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Calcium binding protects E-cadherin from cleavage by Helicobacter pylori HtrA

BACKGROUND: The cell adhesion and tumor suppressor protein E-cadherin is an important factor in the establishment and maintenance of epithelial integrity. E-cadherin is a single transmembrane protein, which consists of an intracellular domain (IC), a transmembrane domain (TD), and five extracellular...

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Autores principales: Schmidt, Thomas P., Goetz, Camilla, Huemer, Markus, Schneider, Gisbert, Wessler, Silja
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4895972/
https://www.ncbi.nlm.nih.gov/pubmed/27274359
http://dx.doi.org/10.1186/s13099-016-0112-6
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author Schmidt, Thomas P.
Goetz, Camilla
Huemer, Markus
Schneider, Gisbert
Wessler, Silja
author_facet Schmidt, Thomas P.
Goetz, Camilla
Huemer, Markus
Schneider, Gisbert
Wessler, Silja
author_sort Schmidt, Thomas P.
collection PubMed
description BACKGROUND: The cell adhesion and tumor suppressor protein E-cadherin is an important factor in the establishment and maintenance of epithelial integrity. E-cadherin is a single transmembrane protein, which consists of an intracellular domain (IC), a transmembrane domain (TD), and five extracellular domains (EC). EC domains form homophilic interactions in cis and trans that require calcium binding to the linker region between the EC domains. In our previous studies, we identified the serine protease high temperature requirement A (HtrA) from the human pathogen and class-I carcinogen Helicobacter pylori (H. pylori) as a bacterial E-cadherin-cleaving protease that targets the linker region of the EC domains, thereby disrupting gastric epithelial integrity. However, it remains unclear how calcium binding to the E-cadherin linker regions affects HtrA-mediated cleavage. RESULTS: Investigating the influence of calcium on the HtrA-mediated cleavage of recombinant E-cadherin (rCdh1) in vitro, we tested different concentrations of calcium ions and the calcium chelator ethylenediaminetetraacetic acid (EDTA). Calcium efficiently reduced HtrA-mediated E-cadherin fragmentation. Conversely, the addition of EDTA strongly increased cleavage, resulting in a ladder of defined E-cadherin fragments. However, calcium ions did not affect HtrA oligomerization and protease activity as monitored by degradation of the universal protease substrate casein. Finally, addition of ethyleneglycol-bis-tetraacetic acid (EGTA) slightly enhanced E-cadherin cleavage during H. pylori infection of gastric epithelial cells. CONCLUSIONS: Our results suggest that calcium blocks HtrA-mediated cleavage by interfering with the accessibility of calcium-binding regions between the individual EC domains, which have been identified as cleavage sites of HtrA. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13099-016-0112-6) contains supplementary material, which is available to authorized users.
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spelling pubmed-48959722016-06-08 Calcium binding protects E-cadherin from cleavage by Helicobacter pylori HtrA Schmidt, Thomas P. Goetz, Camilla Huemer, Markus Schneider, Gisbert Wessler, Silja Gut Pathog Research BACKGROUND: The cell adhesion and tumor suppressor protein E-cadherin is an important factor in the establishment and maintenance of epithelial integrity. E-cadherin is a single transmembrane protein, which consists of an intracellular domain (IC), a transmembrane domain (TD), and five extracellular domains (EC). EC domains form homophilic interactions in cis and trans that require calcium binding to the linker region between the EC domains. In our previous studies, we identified the serine protease high temperature requirement A (HtrA) from the human pathogen and class-I carcinogen Helicobacter pylori (H. pylori) as a bacterial E-cadherin-cleaving protease that targets the linker region of the EC domains, thereby disrupting gastric epithelial integrity. However, it remains unclear how calcium binding to the E-cadherin linker regions affects HtrA-mediated cleavage. RESULTS: Investigating the influence of calcium on the HtrA-mediated cleavage of recombinant E-cadherin (rCdh1) in vitro, we tested different concentrations of calcium ions and the calcium chelator ethylenediaminetetraacetic acid (EDTA). Calcium efficiently reduced HtrA-mediated E-cadherin fragmentation. Conversely, the addition of EDTA strongly increased cleavage, resulting in a ladder of defined E-cadherin fragments. However, calcium ions did not affect HtrA oligomerization and protease activity as monitored by degradation of the universal protease substrate casein. Finally, addition of ethyleneglycol-bis-tetraacetic acid (EGTA) slightly enhanced E-cadherin cleavage during H. pylori infection of gastric epithelial cells. CONCLUSIONS: Our results suggest that calcium blocks HtrA-mediated cleavage by interfering with the accessibility of calcium-binding regions between the individual EC domains, which have been identified as cleavage sites of HtrA. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13099-016-0112-6) contains supplementary material, which is available to authorized users. BioMed Central 2016-06-06 /pmc/articles/PMC4895972/ /pubmed/27274359 http://dx.doi.org/10.1186/s13099-016-0112-6 Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Schmidt, Thomas P.
Goetz, Camilla
Huemer, Markus
Schneider, Gisbert
Wessler, Silja
Calcium binding protects E-cadherin from cleavage by Helicobacter pylori HtrA
title Calcium binding protects E-cadherin from cleavage by Helicobacter pylori HtrA
title_full Calcium binding protects E-cadherin from cleavage by Helicobacter pylori HtrA
title_fullStr Calcium binding protects E-cadherin from cleavage by Helicobacter pylori HtrA
title_full_unstemmed Calcium binding protects E-cadherin from cleavage by Helicobacter pylori HtrA
title_short Calcium binding protects E-cadherin from cleavage by Helicobacter pylori HtrA
title_sort calcium binding protects e-cadherin from cleavage by helicobacter pylori htra
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4895972/
https://www.ncbi.nlm.nih.gov/pubmed/27274359
http://dx.doi.org/10.1186/s13099-016-0112-6
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