The structure of the Shiga toxin 2a A-subunit dictates the interactions of the toxin with blood components.
Animals
Blood Platelets
/ drug effects
Chlorocebus aethiops
Circular Dichroism
Complement Factor H
/ metabolism
Escherichia coli
/ chemistry
Fluorescence
Humans
Leukocytes
/ drug effects
Neutrophils
/ drug effects
Protein Binding
Protein Conformation
Shiga Toxin 2
/ chemistry
Trihexosylceramides
/ metabolism
Trypsin
Vero Cells
Journal
Cellular microbiology
ISSN: 1462-5822
Titre abrégé: Cell Microbiol
Pays: India
ID NLM: 100883691
Informations de publication
Date de publication:
05 2019
05 2019
Historique:
received:
21
09
2018
revised:
06
12
2018
accepted:
17
12
2018
pubmed:
24
12
2018
medline:
31
7
2020
entrez:
23
12
2018
Statut:
ppublish
Résumé
Hemolytic uremic syndrome (eHUS) is a severe complication of human infections with Shiga toxins (Stxs)-producing Escherichia coli. A key step in the pathogenesis of eHUS is the interaction of Stxs with blood components before the targeting of renal endothelial cells. Here, we show that a single proteolytic cleavage in the Stx2a A-subunit, resulting into two fragments (A1 and A2) linked by a disulfide bridge (cleaved Stx2a), dictates different binding abilities. Uncleaved Stx2a was confirmed to bind to human neutrophils and to trigger leukocyte/platelet aggregate formation, whereas cleaved Stx2a was ineffective. Conversely, binding of complement factor H was confirmed for cleaved Stx2a and not for uncleaved Stx2a. It is worth noting that uncleaved and cleaved Stx2a showed no differences in cytotoxicity for Vero cells or Raji cells, structural conformation, and contaminating endotoxin. These results have been obtained by comparing two Stx2a batches, purified in different laboratories by using different protocols, termed Stx2a(cl; cleaved toxin, Innsbruck) and Stx2a(uncl; uncleaved toxin, Bologna). Stx2a(uncl) behaved as Stx2a(cl) after mild trypsin treatment. In this light, previous controversial results obtained with purified Stx2a has to be critically re-evaluated; furthermore, characterisation of the structure of circulating Stx2a is mandatory to understand eHUS-pathogenesis and to develop therapeutic approaches.
Identifiants
pubmed: 30578712
doi: 10.1111/cmi.13000
pmc: PMC6492301
doi:
Substances chimiques
Shiga Toxin 2
0
Trihexosylceramides
0
globotriaosylceramide
71965-57-6
Complement Factor H
80295-65-4
Trypsin
EC 3.4.21.4
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e13000Subventions
Organisme : Austrian Science Fund FWF
ID : W 1253
Pays : Austria
Informations de copyright
© 2018 The Authors Cellular Microbiology Published by John Wiley & Sons Ltd.
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