Colonization factor CS30 from enterotoxigenic Escherichia coli binds to sulfatide in human and porcine small intestine

Astrid Von Mentzer,Dani Zalem,Zofia Chrienova,Susann Teneberg

doi:10.1080/21505594.2020.1749497

Astrid Von Mentzer, Dani Zalem + Show 2 more

Open Access

https://doi.org/10.1080/21505594.2020.1749497

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Abstract

ABSTRACT The ability to adhere via colonization factors to specific receptors located on the intestinal mucosa is a key virulence factor in enterotoxigenic Escherichia coli (ETEC) pathogenesis. Here, the potential glycosphingolipid receptors of the novel human ETEC colonization factor CS30 were examined by binding of CS30-expressing bacteria to glycosphingolipids on thin-layer chromatograms. We thereby found a highly specific binding of CS30-expressing bacteria to a fast-migrating acid glycosphingolipid of human and porcine small intestine, while no binding was obtained with a mutant ETEC strain unable to express CS30 fimbriae. The CS30 binding glycosphingolipid from human small intestine was isolated and characterized by mass spectrometry as sulfatide (SO3-3Galβ1Cer). Comparative binding studies using sulfatides with different ceramide compositions gave a preferential binding of CS30 to sulfatide with d18:1-h24:0 ceramide. This ceramide species of sulfatide was also isolated from human small intestine and characterized by mass spectrometry and antibody binding. These studies implicate sulfatide as candidate receptor for mediating attachment of CS30-fimbriated ETEC to human and porcine small intestinal cells. Our findings may be a basis for designing receptor saccharide analogues for inhibition of the intestinal adhesion of CS30-expressing E. coli.

Highlights

Diarrheal disease is a leading cause of child mortality and morbidity in the world according to the World Health Organization (WHO)
During these studies we repeatedly observed a selective binding of the CS30 expressing enterotoxigenic Escherichia coli (ETEC) to a fastmigrating compound in acid glycosphingolipid fractions (exemplified in Figure 1(b), lanes 1, 3, 4, and 6; Figure 2 (b), lanes 3 and 5)
We investigated the carbohydrate binding specificity of the novel colonization factors (CFs) CS30 by binding of CS30 expressing ETEC to a large number of variant glycosphingolipids

Summary

Introduction

Diarrheal disease is a leading cause of child mortality and morbidity in the world according to the World Health Organization (WHO). The enteric pathogen enterotoxigenic Escherichia coli (ETEC) is the most common cause of bacterial diarrhea in children, mainly in resource-poor regions where access to clean water and proper sanitation are limited [1], and in travelers to endemic areas [2]. Living in close proximity with domestic livestock and poultry is more common in resource-poor countries where animal husbandry serves as a primary source of income. Livestock and domestic animals are common sources of fecal contamination of water and in households [6]. Living with livestock increases the risk of fecal contamination and subsequently elevates the risk of diarrheal pathogen transmission between animals and humans. It has been shown that livestock exposure is associated with diarrheal illness in humans, mainly through fecal contamination of the household environment [7]

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