Casaravilla Cecilia, Díaz Alvaro
Departamento de Biociencias, Facultad de Química and IQB, Universidad de la República, Instituto de Higiene, Montevideo CP, Uruguay.
Mol Biochem Parasitol. 2010 Dec;174(2):132-6. doi: 10.1016/j.molbiopara.2010.07.008. Epub 2010 Aug 6.
The larvae of the cestodes belonging to the genus Echinococcus are outwardly protected by the laminated layer (LL), a crucial but poorly understood carbohydrate-rich acellular structure. Carbohydrate structural data strongly suggest that the main components of the LL are mucins. The most massive LL in the genus is featured by E. granulosus, agent of cystic hydatid disease. No appropriate methods existed to date for the solubilisation of the E. granulosus LL and the electrophoretic visualisation of its proposed structural mucins. We report that reduction of disulphides greatly aids LL disassembly, resulting in almost full solubilisation in combination with moderate sonication. The structural mucins can then be visualised by agarose electrophoresis and blotting with galactose-binding lectins, which also react strongly with the LL in tissue sections. A substantial portion of the material migrates as if positively charged; since the LL glycans are neutral, this may correspond to mucins with cationic peptide backbones.
属于棘球绦虫属的绦虫幼虫在外部由分层层(LL)保护,这是一种关键但了解甚少的富含碳水化合物的无细胞结构。碳水化合物结构数据有力地表明,LL的主要成分是粘蛋白。该属中最厚的LL以细粒棘球绦虫为特征,它是囊型包虫病的病原体。迄今为止,尚无合适的方法来溶解细粒棘球绦虫的LL并对其推测的结构粘蛋白进行电泳可视化。我们报告说,二硫键的还原极大地有助于LL的拆解,与适度超声处理相结合可实现几乎完全溶解。然后可以通过琼脂糖电泳和用半乳糖结合凝集素进行印迹来观察结构粘蛋白,这些凝集素在组织切片中也与LL发生强烈反应。相当一部分物质迁移时似乎带正电;由于LL聚糖是中性的,这可能对应于具有阳离子肽主链的粘蛋白。