Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Enzymatic Cleavage of Stx2a in the Gut and Identification of Pancreatic Elastase and Trypsin as Possible Main Cleavers

Sára Kellnerová
,
Silke Huber
ORCID logo ,
Mariam Massri
ORCID logo ,
Verena Fleischer
,
Klemens Losso
ORCID logo ,
Bettina Sarg
,
Leopold Kremser
,
Heribert Talasz
ORCID logo ,
Xiaohua He
,
Elisa Varrone
,
Maurizio Brigotti
,
Gianluigi Ardissino
,
Dorothea Orth-Höller
* ORCID logo ,
Reinhard Würzner
*
Version 1 : Received: 6 July 2023 / Approved: 7 July 2023 / Online: 7 July 2023 (10:24:04 CEST)

A peer-reviewed article of this Preprint also exists.

Kellnerová, S.; Huber, S.; Massri, M.; Fleischer, V.; Losso, K.; Sarg, B.; Kremser, L.; Talasz, H.; He, X.; Varrone, E.; Brigotti, M.; Ardissino, G.; Orth-Höller, D.; Würzner, R. Enzymatic Cleavage of Stx2a in the Gut and Identification of Pancreatic Elastase and Trypsin as Possible Main Cleavers. Microorganisms 2023, 11, 2487. Kellnerová, S.; Huber, S.; Massri, M.; Fleischer, V.; Losso, K.; Sarg, B.; Kremser, L.; Talasz, H.; He, X.; Varrone, E.; Brigotti, M.; Ardissino, G.; Orth-Höller, D.; Würzner, R. Enzymatic Cleavage of Stx2a in the Gut and Identification of Pancreatic Elastase and Trypsin as Possible Main Cleavers. Microorganisms 2023, 11, 2487.

Abstract

Shiga toxins (Stxs), and more specifically 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. Cleavage of Stxs A subunit, when followed by reduction, boosts the enzymatic activity causing damage to targeted cells. This cleavage was assumed to be mostly mediated by furin during Stx intracellular trafficking. Cleavage of A subunit, without reduction, changes the binding properties of Stxs for blood components during HUS pathogenesis. To investigate whether the cleavage could occur in the intestine, even prior entering target cells, Stx2a structure was characterized after its exposure to specific host factors present in human stool. The structure of Stx2a A subunit was determined by immunoblotting after electrophoretic separation under reducing conditions. Trypsin and chymotrypsin-like elastase 3B (CELA3B), two serine proteases, were identified as potential candidates that may trigger the extracellular cleavage of Stx2a A subunit directly after its secretion by EHEC in the gut. Whether this observation indeed translates to natural infections and plays a role in eHUS pathogenesis has yet to be determined as well as whether host's protease profile could affect disease development by changing the toxin’s biological features.

Keywords

enterohemorrhagic Escherichia coli (EHEC); EHEC-associated hemolytic uremic syndrome (eHUS); Shiga toxin 2a (Stx2a); trypsin; furin; chymotrypsin-like elastase 3B (CELA3B)

Subject

Biology and Life Sciences, Immunology and Microbiology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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