Functional Analysis of Botulinum Hemagglutinin (HA)

Methods Mol Biol. 2020;2132:191-200. doi: 10.1007/978-1-0716-0430-4_20.


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.

Keywords: Botulinum; Carbohydrate-binding activity; HA; Hemagglutinin; Intestinal absorption; Lectin activity; Microfold cell; Neurotoxin; Transcytosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adsorption
  • Animals
  • Botulinum Toxins / metabolism
  • Caco-2 Cells
  • Cadherins / chemistry
  • Cadherins / metabolism*
  • Cell Line
  • Clostridium botulinum / genetics
  • Clostridium botulinum / metabolism*
  • Dogs
  • Epithelial Cells / metabolism
  • Epithelial Cells / microbiology
  • Female
  • Foodborne Diseases / microbiology
  • Hemagglutinins / chemistry
  • Hemagglutinins / genetics
  • Hemagglutinins / pharmacology*
  • Humans
  • Intestinal Mucosa / metabolism*
  • Intestinal Mucosa / microbiology
  • Madin Darby Canine Kidney Cells
  • Mice
  • Protein Binding / drug effects
  • Protein Engineering


  • Cadherins
  • Cdh1 protein, mouse
  • Hemagglutinins
  • Botulinum Toxins