Abstract
Botulinum neurotoxin (BoNT), produced by Clostridium botulinum, is the most potent toxin and produced as a complex with non-toxic components. Food-borne botulism is caused by the ingestion of these BoNT complexes. Hemagglutinin (HA), one of the non-toxic components, is known to have lectin (carbohydrate binding) activity and E-cadherin-binding activity. These activities promote the intestinal absorption of BoNT. To elucidate the mechanism of the onset of food-borne botulism, we focused on the role of HA in the intestinal absorption of BoNT. We describe the functional analysis methods for HA, including the expression of recombinant proteins, binding to glycoproteins and epithelial cells, and localization in mouse intestinal tissue.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Schiavo G, Matteoli M, Montecucco C (2000) Neurotoxins affecting neuroexocytosis. Physiol Rev 80:717–766
Rossetto O, Pirazzini M, Montecucco C (2014) Botulinum neurotoxins: genetic, structural and mechanistic insights. Nat Rev Microbial 12:535–549
Sakaguchi G (1982) Clostridium botulinum toxins. Pharmacol Ther 19:165–194
Fujinaga Y, Sugawara Y, Matsumura T (2013) Uptake of botulinum neurotoxin in the intestine. Curr Top Microbiol Immunol 364:45–59
Gu S, Rumpel S, Zhou J, Strotmeier J, Bigalke H et al (2012) Botulinum neurotoxin is shielded by NTNHA in an interlocked complex. Science 335:977–981
Benefield DA, Dessain SK, Shine N, Ohi MD, Lacy DB (2013) Molecular assembly of botulinum neurotoxin progenitor complexes. Proc Natl Acad Sci U S A 110:5630–5635
Lee K, Gu S, Jin L, Le TT CLW et al (2013) Structure of a bimodular botulinum neurotoxin complex provides insights into its oral toxicity. PLoS Pathog 9:e1003690
Amatsu S, Sugawara Y, Matsumura T, Kitadokoro K, Fujinaga Y (2013) Crystal structure of Clostridium botulinum whole hemagglutinin reveals a huge triskelion-shaped molecular complex. J Biol Chem 288:35617–35625
Fujinaga Y, Inoue K, Watanabe S et al (1997) The haemagglutinin of Clostridium botulinum type C progenitor toxin plays an essential role in binding of toxin to the epithelial cells of Guinea pig small intestine, leading to the efficient absorption of the toxin. Microbiology 143:3841–3847
Fujinaga Y, Inoue K, Nomura T et al (2000) Identification and characterization of functional subunits of Clostridium botulinum type A progenitor toxin involved in binding to intestinal microvilli and erythrocytes. FEBS Lett 467, 179:–83
Fujinaga Y, Inoue K, Watarai S et al (2004) Molecular characterization of binding subcomponents of Clostridium botulinum type C progenitor toxin for intestinal epithelial cells and erythrocytes. Microbiology 150:1529–1538
Matsumura T, Jin Y, Kabumoto Y et al (2008) The HA proteins of botulinum toxin disrupt intestinal epithelial intercellular junctions to increase toxin absorption. Cell Microbiol 10:355–364
Jin Y, Takegahara Y, Sugawara Y et al (2009) Disruption of the epithelial barrier by botulinum haemagglutinin (HA) proteins - differences in cell tropism and the mechanism of action between HA proteins of types A or B, and HA proteins of type C. Microbiology 155:35–45
Sugawara Y, Matsumura T, Takegahara Y et al (2010) Botulinum hemagglutinin disrupts the intercellular epithelial barrier by directly binding E-cadherin. J Cell Biol 189:691–700
Sugawara Y, Yutani M, Amatsu S et al (2014) Functional dissection of the Clostridium botulinum type B hemagglutinin complex: identification of the carbohydrate and E-cadherin binding sites. PLoS One 9:e111170
Lee K, Zhong X, Gu S, Kruel AM, Dorner MB et al (2014) Molecular basis for disruption of E-cadherin adhesion by botulinum neurotoxin a complex. Science 344:1405–1410
Matsumura T, Sugawara Y, Yutani M, Amatsu S, Yagita H et al (2015) Botulinum toxin a complex exploits intestinal M cells to enter the host and exert neurotoxicity. Nat Commun \: 6255
Amatsu S, Matsumura T, Yutani M, Fujinaga Y (2018) Multivalency effects of hemagglutinin component of type B botulinum neurotoxin complex on epithelial barrier disruption. Microbiol Immunol 62:80–89
Acknowledgments
This study was supported in part by grants to T.M. from the Japan Society for the Promotion of Science KAKENHI (Grant No. 16Â K19123, 18Â K07107), the Takeda Science Foundation, SUZUKEN Memorial Foundation, the Sekisui Chemical Grant Program, and grants to Y.F. from the Promotion of Science KAKENHI (Grant No. 18H02654)Â and the Japanese Initiative for Progress of Research on Infectious Disease for global Epidemic (J-PRIDE, Grant No. JP18fm0208010).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Matsumura, T., Fujinaga, Y. (2020). Functional Analysis of Botulinum Hemagglutinin (HA). In: Hirabayashi, J. (eds) Lectin Purification and Analysis. Methods in Molecular Biology, vol 2132. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0430-4_20
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0430-4_20
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0429-8
Online ISBN: 978-1-0716-0430-4
eBook Packages: Springer Protocols