Immunocytochemical Detection of Recombinant Biomphalysin on Schistosoma mansoni Sporocysts

Abstract

Schistosomiasis, or bilharzia, is a tropical disease caused by worms of the genus Schistosoma which infect about 200 million people. The life cycle of the parasite requires Biomphalaria, a specific genus of freshwater snails, as intermediate. Using an interactome approach employing B. glabrata plasma and S. mansoni primary sporocyst extracts, we identified a new cytolytic protein called Biomphalysin that displays similarities to members of the β-PFT superfamily known to form channels in targeted membranes. To investigate its mechanism of action, we produced a recombinant protein flanked by an N-terminal 6 histidine tag. Then, we investigated the ability of Biomphalysin to interact with the sporocyst tegument. This optimized protocol describes an immunocytochemical procedure to detect histidine tagged recombinant protein on the sporocyst tegumental membrane., 血吸虫病或鼠伤寒沙菌是由感染约2亿人的蠕虫属引起的热带病。 寄生虫的生命周期需要作为中间体的淡水蜗牛的特定属的生物疟原虫。 使用采用em的interactome方法。 glabrata plasma和 我们鉴定了一种称为生物溶解素的新的溶细胞蛋白，其显示与已知在靶膜中形成通道的β-PFT超家族成员的相似性。 为了调查其作用机制，我们产生侧翼有N端6组氨酸标签的重组蛋白。 然后，我们调查了Biomphalysin与孢子囊体积相互作用的能力。 这个优化的协议描述免疫细胞化学程序检测组氨酸标记的重组蛋白在孢子被膜上。