Cargando…
Enzymatic Cleavage of Stx2a in the Gut and Identification of Pancreatic Elastase and Trypsin as Possible Main Cleavers
Shiga toxins (Stxs), especially the Stx2a subtype, are the major virulence factors involved in enterohemorrhagic Escherichia coli (EHEC)-associated hemolytic uremic syndrome (eHUS), a life-threatening disease causing acute kidney injury, especially in children. After oral transmission and colonizati...
Autores principales: | , , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10609011/ https://www.ncbi.nlm.nih.gov/pubmed/37894145 http://dx.doi.org/10.3390/microorganisms11102487 |
_version_ | 1785127913263202304 |
---|---|
author | Kellnerová, Sára Huber, Silke Massri, Mariam Fleischer, Verena Losso, Klemens Sarg, Bettina Kremser, Leopold Talasz, Heribert He, Xiaohua Varrone, Elisa Brigotti, Maurizio Ardissino, Gianluigi Orth-Höller, Dorothea Würzner, Reinhard |
author_facet | Kellnerová, Sára Huber, Silke Massri, Mariam Fleischer, Verena Losso, Klemens Sarg, Bettina Kremser, Leopold Talasz, Heribert He, Xiaohua Varrone, Elisa Brigotti, Maurizio Ardissino, Gianluigi Orth-Höller, Dorothea Würzner, Reinhard |
author_sort | Kellnerová, Sára |
collection | PubMed |
description | Shiga toxins (Stxs), especially the Stx2a subtype, are the major virulence factors involved in enterohemorrhagic Escherichia coli (EHEC)-associated hemolytic uremic syndrome (eHUS), a life-threatening disease causing acute kidney injury, especially in children. After oral transmission and colonization in the gut, EHEC release Stx. Intracellular cleavage of the Stx A subunit, when followed by reduction, boosts the enzymatic activity that causes damage to targeted cells. This cleavage was assumed to be mostly mediated by furin during Stx intracellular trafficking. To investigate whether this cleavage could occur in the intestine, even prior to entering target cells, Stx2a A subunit structure (intact or cleaved) was characterized after its exposure to specific host factors present in human stool. The molecular weight of Stx2a A subunit/fragments was determined by immunoblotting after electrophoretic separation under reducing conditions. In this study, it was demonstrated that Stx2a is cleaved by certain human stool components. Trypsin and chymotrypsin-like elastase 3B (CELA3B), two serine proteases, were identified as potential candidates that can trigger the extracellular cleavage of Stx2a A subunit directly after its secretion by EHEC in the gut. Whether the observed cleavage indeed translates to natural infections and plays a role in eHUS pathogenesis has yet to be determined. If so, it seems likely that a host’s protease profile could affect disease development by changing the toxin’s biological features. |
format | Online Article Text |
id | pubmed-10609011 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-106090112023-10-28 Enzymatic Cleavage of Stx2a in the Gut and Identification of Pancreatic Elastase and Trypsin as Possible Main Cleavers Kellnerová, Sára Huber, Silke Massri, Mariam Fleischer, Verena Losso, Klemens Sarg, Bettina Kremser, Leopold Talasz, Heribert He, Xiaohua Varrone, Elisa Brigotti, Maurizio Ardissino, Gianluigi Orth-Höller, Dorothea Würzner, Reinhard Microorganisms Article Shiga toxins (Stxs), especially the Stx2a subtype, are the major virulence factors involved in enterohemorrhagic Escherichia coli (EHEC)-associated hemolytic uremic syndrome (eHUS), a life-threatening disease causing acute kidney injury, especially in children. After oral transmission and colonization in the gut, EHEC release Stx. Intracellular cleavage of the Stx A subunit, when followed by reduction, boosts the enzymatic activity that causes damage to targeted cells. This cleavage was assumed to be mostly mediated by furin during Stx intracellular trafficking. To investigate whether this cleavage could occur in the intestine, even prior to entering target cells, Stx2a A subunit structure (intact or cleaved) was characterized after its exposure to specific host factors present in human stool. The molecular weight of Stx2a A subunit/fragments was determined by immunoblotting after electrophoretic separation under reducing conditions. In this study, it was demonstrated that Stx2a is cleaved by certain human stool components. Trypsin and chymotrypsin-like elastase 3B (CELA3B), two serine proteases, were identified as potential candidates that can trigger the extracellular cleavage of Stx2a A subunit directly after its secretion by EHEC in the gut. Whether the observed cleavage indeed translates to natural infections and plays a role in eHUS pathogenesis has yet to be determined. If so, it seems likely that a host’s protease profile could affect disease development by changing the toxin’s biological features. MDPI 2023-10-04 /pmc/articles/PMC10609011/ /pubmed/37894145 http://dx.doi.org/10.3390/microorganisms11102487 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Kellnerová, Sára Huber, Silke Massri, Mariam Fleischer, Verena Losso, Klemens Sarg, Bettina Kremser, Leopold Talasz, Heribert He, Xiaohua Varrone, Elisa Brigotti, Maurizio Ardissino, Gianluigi Orth-Höller, Dorothea Würzner, Reinhard Enzymatic Cleavage of Stx2a in the Gut and Identification of Pancreatic Elastase and Trypsin as Possible Main Cleavers |
title | Enzymatic Cleavage of Stx2a in the Gut and Identification of Pancreatic Elastase and Trypsin as Possible Main Cleavers |
title_full | Enzymatic Cleavage of Stx2a in the Gut and Identification of Pancreatic Elastase and Trypsin as Possible Main Cleavers |
title_fullStr | Enzymatic Cleavage of Stx2a in the Gut and Identification of Pancreatic Elastase and Trypsin as Possible Main Cleavers |
title_full_unstemmed | Enzymatic Cleavage of Stx2a in the Gut and Identification of Pancreatic Elastase and Trypsin as Possible Main Cleavers |
title_short | Enzymatic Cleavage of Stx2a in the Gut and Identification of Pancreatic Elastase and Trypsin as Possible Main Cleavers |
title_sort | enzymatic cleavage of stx2a in the gut and identification of pancreatic elastase and trypsin as possible main cleavers |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10609011/ https://www.ncbi.nlm.nih.gov/pubmed/37894145 http://dx.doi.org/10.3390/microorganisms11102487 |
work_keys_str_mv | AT kellnerovasara enzymaticcleavageofstx2ainthegutandidentificationofpancreaticelastaseandtrypsinaspossiblemaincleavers AT hubersilke enzymaticcleavageofstx2ainthegutandidentificationofpancreaticelastaseandtrypsinaspossiblemaincleavers AT massrimariam enzymaticcleavageofstx2ainthegutandidentificationofpancreaticelastaseandtrypsinaspossiblemaincleavers AT fleischerverena enzymaticcleavageofstx2ainthegutandidentificationofpancreaticelastaseandtrypsinaspossiblemaincleavers AT lossoklemens enzymaticcleavageofstx2ainthegutandidentificationofpancreaticelastaseandtrypsinaspossiblemaincleavers AT sargbettina enzymaticcleavageofstx2ainthegutandidentificationofpancreaticelastaseandtrypsinaspossiblemaincleavers AT kremserleopold enzymaticcleavageofstx2ainthegutandidentificationofpancreaticelastaseandtrypsinaspossiblemaincleavers AT talaszheribert enzymaticcleavageofstx2ainthegutandidentificationofpancreaticelastaseandtrypsinaspossiblemaincleavers AT hexiaohua enzymaticcleavageofstx2ainthegutandidentificationofpancreaticelastaseandtrypsinaspossiblemaincleavers AT varroneelisa enzymaticcleavageofstx2ainthegutandidentificationofpancreaticelastaseandtrypsinaspossiblemaincleavers AT brigottimaurizio enzymaticcleavageofstx2ainthegutandidentificationofpancreaticelastaseandtrypsinaspossiblemaincleavers AT ardissinogianluigi enzymaticcleavageofstx2ainthegutandidentificationofpancreaticelastaseandtrypsinaspossiblemaincleavers AT orthhollerdorothea enzymaticcleavageofstx2ainthegutandidentificationofpancreaticelastaseandtrypsinaspossiblemaincleavers AT wurznerreinhard enzymaticcleavageofstx2ainthegutandidentificationofpancreaticelastaseandtrypsinaspossiblemaincleavers |