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Bovine Muc1 inhibits binding of enteric bacteria to Caco-2 cells

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Abstract

Inhibition of bacterial adhesion to intestinal epithelial receptors by the consumption of natural food components is an attractive strategy for the prevention of microbial related gastrointestinal illness. We hypothesised that Muc1, a highly glycosylated mucin present in cows’ milk, may be one such food component. Purified bovine Muc1 was tested for its ability to inhibit binding of common enteric bacterial pathogens to Caco-2 cells grown in vitro. Muc1 caused dose-dependent binding inhibition of Escherichia coli, Salmonella enterica serovar Typhimurium (S. Typhimurium), Staphylococcus aureus and Bacillus subtilis. This inhibition was more pronounced for the Gram negative compared with Gram positive bacteria. It was also demonstrated that Muc1, immobilised on a membrane, bound all these bacterial species in a dose-dependent manner, although there was greater interaction with the Gram negative bacteria. A range of monosaccharides, representative of the Muc1 oligosaccharide composition, were tested for their ability to prevent binding of E. coli and S. Typhimurium to Caco-2 cells. Inhibition was structure dependent with sialic acid, L(-) fucose and D(+) mannose significantly inhibiting binding of both Gram negative species. N-acetylglucosamine and N-acetylgalactosamine significantly inhibited binding of E. coli whilst galactose, one of the most abundant Muc1 monosaccharides, showed the strongest inhibition against S. Typhimurium. Treatment with sialidase significantly decreased the inhibitory properties of Muc1, demonstrating the importance of sialic acid in adhesion inhibition. It is concluded that bovine Muc1 prevents binding of bacteria to human intestinal cells and may have a role in preventing the binding of common enteropathogenic bacteria to human intestinal epithelial surfaces.

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Acknowledgements

This research was supported by the Geoffrey Gardiner Dairy Foundation and the Co-operative Research Centre for Innovative Dairy Products. Access to the Australian Proteome Analysis Facility was aided by the Australian Government’s Major National Research Facilities Program. The authors thank Parmalat Australia Ltd. for their donation of cream for isolation of Muc1.

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Authors and Affiliations

  1. Faculty of Health, Medicine, Nursing and Behavioural Sciences, School of Exercise and Nutrition Sciences, Deakin University, 221 Burwood Highway, Burwood, 3125, Victoria, Australia

    Phillip Parker & Stuart Smith

  2. CSIRO Livestock Industries, St Lucia, Queensland, 4067, Australia

    Lillian Sando, Roger Pearson, Kritaya Kongsuwan & Ross L. Tellam

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  1. Phillip Parker
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  2. Lillian Sando
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  3. Roger Pearson
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Correspondence to Phillip Parker.

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Author’s contributions

PP and LS contributed equally to this research. PP carried out all the direct binding and inhibition assays and monosaccharide binding studies. LS and RP carried out the protein purification, SDS-PAGE analysis and sialidase treatment. RT, SS and KK participated in the conception, design and overall coordination of the study and provided advice on the various assays. All authors have read and approved the final manuscript.

Phillip Parker and Lillian Sando contributed equally to this research.

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Parker, P., Sando, L., Pearson, R. et al. Bovine Muc1 inhibits binding of enteric bacteria to Caco-2 cells. Glycoconj J 27, 89–97 (2010). https://doi.org/10.1007/s10719-009-9269-2

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  • DOI: https://doi.org/10.1007/s10719-009-9269-2

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