McDiarmid S S, Dean L L, Podesta R B
Mol Biochem Parasitol. 1983 Feb;7(2):141-57. doi: 10.1016/0166-6851(83)90041-5.
The outer and inner bilayers of the apical membrane complex of Schistosoma mansoni were sequentially stripped from adult worms by two incubations in 0.1% digitonin solutions. Membrane removal was evaluated by electron microscopy of worms and bilayer material, using Con A-ferritin as a marker for the outer bilayer. Amounts of Con A removed by the digests were measured with a tritiated Con A marker. To measure the purity of the fractions membrane markers were characterised and quantitated for both bilayers. In the absence of the usual enzymatic markers for plasma membrane diazotised [125I]-iodosulfanilic acid was used as a marker for the outer bilayer. Alkaline phosphatase and a Na+, Mg2+-ATPase were localised to the inner bilayer. From these results we can deduce that the inner bilayer is analogous to the typical, apical plasma membrane of other animal epithelia. The outer bilayer does not share these enzymatic similarities. The integrity of the syncytium after removal of the outer bilayer and the increased levels of lactate dehydrogenase in the supernatant after removal of the inner bilayer suggests that the outer bilayer is secondary in maintaining the permeability barrier of the apical membrane complex, with respect to soluble proteins. The possible significance of these results in terms of the destructive action of complement on the parasite are discussed.
通过在0.1%洋地黄皂苷溶液中进行两次孵育,依次从曼氏血吸虫成虫中剥离出顶膜复合体的外层和内层双层膜。通过对虫体和双层膜材料进行电子显微镜观察,并使用伴刀豆球蛋白A-铁蛋白作为外层双层膜的标记物,来评估膜的去除情况。用氚化伴刀豆球蛋白A标记物测量消化物中去除的伴刀豆球蛋白A的量。为了测量各组分的纯度,对双层膜的膜标记物进行了表征和定量分析。在缺乏质膜常用酶标记物的情况下,用重氮化的[125I]-碘磺胺酸作为外层双层膜的标记物。碱性磷酸酶和Na+、Mg2+-ATP酶定位于内层双层膜。从这些结果我们可以推断,内层双层膜类似于其他动物上皮细胞典型的顶端质膜。外层双层膜不具有这些酶学相似性。去除外层双层膜后合体细胞的完整性以及去除内层双层膜后上清液中乳酸脱氢酶水平的升高表明,就可溶性蛋白质而言,外层双层膜在维持顶膜复合体的通透性屏障方面是次要的。讨论了这些结果在补体对寄生虫破坏作用方面的可能意义。