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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...

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Autores principales: 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
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
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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.
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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
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