Abstract
Studies of the interaction of bacteria with mucus-secreting cells can be complemented at a more mechanistic level by exploring the interaction of bacteria with purified mucins. Here we describe a far Western blotting approach to show how C. jejuni proteins separated by SDS PAGE and transferred to a membrane or slot blotted directly onto a membrane can be probed using biotinylated mucin. In addition we describe the use of novel mucin microarrays to assess bacterial interactions with mucins in a high-throughput manner.
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References
Naughton JA, Marino K, Dolan B et al (2013) Divergent mechanisms of interaction of Helicobacter pylori and Campylobacter jejuni with mucus and mucins. Infect Immun 81(8):2838–2850. doi:10.1128/IAI.00415-13
Tu QV, McGuckin MA, Mendz GL (2008) Campylobacter jejuni response to human mucin MUC2: modulation of colonization and pathogenicity determinants. J Med Microbiol 57(Pt 7):795–802. doi:10.1099/jmm.0.47752-0
Alemka A, Whelan S, Gough R et al (2010) Purified chicken intestinal mucin attenuates Campylobacter jejuni pathogenicity in vitro. J Med Microbiol 59(Pt 8):898–903. doi:10.1099/jmm.0.019315-0
Byrne CM, Clyne M, Bourke B (2007) Campylobacter jejuni adhere to and invade chicken intestinal epithelial cells in vitro. Microbiology 153(Pt 2):561–569. doi:10.1099/mic.0.2006/000711-0
McAuley JL, Linden SK, Png CW et al (2007) MUC1 cell surface mucin is a critical element of the mucosal barrier to infection. J Clin Invest 117(8):2313–2324. doi:10.1172/JCI26705
Linden SK, Florin TH, McGuckin MA (2008) Mucin dynamics in intestinal bacterial infection. PLoS One 3(12), e3952. doi:10.1371/journal.pone.0003952
Dawson PA, Huxley S, Gardiner B et al (2009) Reduced mucin sulfonation and impaired intestinal barrier function in the hyposulfataemic NaS1 null mouse. Gut 58(7):910–919. doi:10.1136/gut.2007.147595
Kilcoyne M, Gerlach JQ, Gough R et al (2012) Construction of a natural mucin microarray and interrogation for biologically relevant glyco-epitopes. Anal Chem 84(7):3330–3338. doi:10.1021/ac203404n
Acknowledgments
This work was supported by a grant from Science Foundation Ireland (08/SRC/B1393).
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Clyne, M., Duggan, G., Naughton, J., Bourke, B. (2017). Methods to Assess the Direct Interaction of C. jejuni with Mucins. In: Butcher, J., Stintzi, A. (eds) Campylobacter jejuni. Methods in Molecular Biology, vol 1512. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6536-6_10
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DOI: https://doi.org/10.1007/978-1-4939-6536-6_10
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-6536-6
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