Structure of lymphostatin, a large multi-functional virulence factor of pathogenic Escherichia coli

Abstract Lymphostatin is a key virulence factor of enteropathogenic and enterohaemorrhagic Escherichia coli, playing roles in bacterial colonisation of the gut and in the inhibition of lymphocyte proliferation and proinflammatory responses. The protein’s glycosyltransferase and cysteine protease mot...

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Main Authors: Matthias Griessmann, Tim Rasmussen, Vanessa J. Flegler, Christian Kraft, Ronja Schneider, Max Hateley, Lukas Spantzel, Mark P. Stevens, Michael Börsch, Bettina Böttcher
Format: Article
Language:English
Published: Nature Portfolio 2025-06-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-60995-9
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author Matthias Griessmann
Tim Rasmussen
Vanessa J. Flegler
Christian Kraft
Ronja Schneider
Max Hateley
Lukas Spantzel
Mark P. Stevens
Michael Börsch
Bettina Böttcher
author_facet Matthias Griessmann
Tim Rasmussen
Vanessa J. Flegler
Christian Kraft
Ronja Schneider
Max Hateley
Lukas Spantzel
Mark P. Stevens
Michael Börsch
Bettina Böttcher
author_sort Matthias Griessmann
collection DOAJ
description Abstract Lymphostatin is a key virulence factor of enteropathogenic and enterohaemorrhagic Escherichia coli, playing roles in bacterial colonisation of the gut and in the inhibition of lymphocyte proliferation and proinflammatory responses. The protein’s glycosyltransferase and cysteine protease motifs are required for activity against lymphocytes, but high-resolution structural information has proven elusive. Here, we describe the structure of lymphostatin from enteropathogenic E. coli O127:H6, determined by electron cryo-microscopy at different pH values. We observe three conformations of a highly complex molecule with two glycosyltransferase domains, one PaToxP-like protease domain, an ADP-ribosyltransferase domain, a vertex domain and a delivery domain. Long linkers hold these domains together and occlude the catalytic sites of the N-terminal glycosyltransferase and protease domains. Lymphostatin binds to bovine T-lymphocytes and HEK-293T cells, forming clusters at the plasma membrane that are internalized. With six distinct domains, lymphostatin can be regarded as a multitool of pathogenic Escherichia coli, enabling complex interactions with host cells.
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spelling doaj-art-bd0ac145035a41d181643e79bdb82aec2025-08-20T03:27:18ZengNature PortfolioNature Communications2041-17232025-06-0116111410.1038/s41467-025-60995-9Structure of lymphostatin, a large multi-functional virulence factor of pathogenic Escherichia coliMatthias Griessmann0Tim Rasmussen1Vanessa J. Flegler2Christian Kraft3Ronja Schneider4Max Hateley5Lukas Spantzel6Mark P. Stevens7Michael Börsch8Bettina Böttcher9University of Würzburg, Rudolf Virchow Center and BiocenterUniversity of Würzburg, Rudolf Virchow Center and BiocenterUniversity of Würzburg, Rudolf Virchow Center and BiocenterUniversity of Würzburg, Rudolf Virchow Center and BiocenterUniversity of Würzburg, Rudolf Virchow Center and BiocenterUniversity of Edinburgh, The Roslin Institute and Royal (Dick) School of Veterinary StudiesJena University Hospital, Single Molecule Microscopy GroupUniversity of Edinburgh, The Roslin Institute and Royal (Dick) School of Veterinary StudiesJena University Hospital, Single Molecule Microscopy GroupUniversity of Würzburg, Rudolf Virchow Center and BiocenterAbstract Lymphostatin is a key virulence factor of enteropathogenic and enterohaemorrhagic Escherichia coli, playing roles in bacterial colonisation of the gut and in the inhibition of lymphocyte proliferation and proinflammatory responses. The protein’s glycosyltransferase and cysteine protease motifs are required for activity against lymphocytes, but high-resolution structural information has proven elusive. Here, we describe the structure of lymphostatin from enteropathogenic E. coli O127:H6, determined by electron cryo-microscopy at different pH values. We observe three conformations of a highly complex molecule with two glycosyltransferase domains, one PaToxP-like protease domain, an ADP-ribosyltransferase domain, a vertex domain and a delivery domain. Long linkers hold these domains together and occlude the catalytic sites of the N-terminal glycosyltransferase and protease domains. Lymphostatin binds to bovine T-lymphocytes and HEK-293T cells, forming clusters at the plasma membrane that are internalized. With six distinct domains, lymphostatin can be regarded as a multitool of pathogenic Escherichia coli, enabling complex interactions with host cells.https://doi.org/10.1038/s41467-025-60995-9
spellingShingle Matthias Griessmann
Tim Rasmussen
Vanessa J. Flegler
Christian Kraft
Ronja Schneider
Max Hateley
Lukas Spantzel
Mark P. Stevens
Michael Börsch
Bettina Böttcher
Structure of lymphostatin, a large multi-functional virulence factor of pathogenic Escherichia coli
Nature Communications
title Structure of lymphostatin, a large multi-functional virulence factor of pathogenic Escherichia coli
title_full Structure of lymphostatin, a large multi-functional virulence factor of pathogenic Escherichia coli
title_fullStr Structure of lymphostatin, a large multi-functional virulence factor of pathogenic Escherichia coli
title_full_unstemmed Structure of lymphostatin, a large multi-functional virulence factor of pathogenic Escherichia coli
title_short Structure of lymphostatin, a large multi-functional virulence factor of pathogenic Escherichia coli
title_sort structure of lymphostatin a large multi functional virulence factor of pathogenic escherichia coli
url https://doi.org/10.1038/s41467-025-60995-9
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