Studies on the structural mucins of the Echinococcus granulosus laminated layer

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.