Department of Bacteriology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan.
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.
肉毒神经毒素(BoNT)由肉毒梭菌产生,是最有效的毒素,以复合物的形式与无毒成分共同存在。食源性肉毒中毒是由摄入这些 BoNT 复合物引起的。血凝素(HA)是一种无毒成分,已知具有凝集素(碳水化合物结合)活性和 E-钙黏蛋白结合活性。这些活性促进了 BoNT 的肠道吸收。为了阐明食源性肉毒中毒的发病机制,我们专注于 HA 在 BoNT 肠道吸收中的作用。我们描述了 HA 的功能分析方法,包括重组蛋白的表达、与糖蛋白和上皮细胞的结合以及在小鼠肠道组织中的定位。