The secretome of a parasite in its definitive host can be considered to be its genome in trans, to the extent that secreted products encoded by the parasite fulfill their function in the host milieu. The 'extended phenotype' of the filarial parasite, Brugia malayi, is of particular interest because of the evidence that infection results in potent down-modulation of the host immune response. We collected B. malayi 'excretory-secretory' (BES) proteins from adult parasites and using a combination of shotgun LC-MS/MS and 2D gel electrophoresis, identified 80 B. malayi and two host proteins in BES, of which 31 (38%) were detectable in whole worm extract (BmA). Products which were enriched in BES relative to BmA included phosphatidylethanolamine-binding protein (PEB), leucyl aminopeptidase (LAP, homologue of ES-62 from the related filaria Acanthocheilonema viteae), N-acetylglucosaminyltransferase (GlcNAcT) and galectin-1, in addition to the previously described major surface glycoprotein, glutathione peroxidase (gp29, GPX-1) and the cytokine homologue macrophage migration inhibitory factor (MIF-1). One of the most abundant released proteins was triose phosphate isomerase (TPI), yet many other glycolytic enzymes (such as aldolase and GAPDH) were found only in the somatic extract. Among the more prominent novel products identified in BES were a set of 11 small transthyretin-like proteins, and three glutamine-rich-repeat mucin-like proteins. Notably, no evidence was found of any secreted protein corresponding to the genome of the Wolbachia endosymbiont present in B. malayi. Western blotting with anti-phosphorylcholine (PC) monoclonal antibody identified that GlcNAcT, and not the ES-62 homologue, is the major PC-bearing protein in BES, while probing with human filariasis sera showed preferential reactivity to galectin-1 and to processed forms of myotactin. Overall, this analysis demonstrates selective release of a suite of newly identified proteins not previously suspected to be involved at the host-parasite interface, and provides important new perspectives on the biology of the filarial parasite.

译文

:在寄生虫编码的分泌产物在宿主环境中发挥其功能的范围内,其最终宿主中的寄生虫的分泌组可以认为是其反基因组。丝状寄生虫马来氏布鲁氏菌的“扩展表型”特别受关注,因为有证据表明感染会导致宿主免疫反应强烈下调。我们从成虫体内收集了马来芽孢杆菌的“排泄分泌物”(BES)蛋白,并使用of弹枪LC-MS / MS和2D凝胶电泳的组合,在BES中鉴定出80个马来芽孢杆菌和两种宿主蛋白​​,其中31(38 %)在整个蠕虫提取物(BmA)中均可检测到。相对于BmA而言,富含BES的产品还包括磷脂酰乙醇胺结合蛋白(PEB),亮氨酰肽酶(LAP,来自相关丝虫棘皮动物丝虫的ES-62的同源物),N-乙酰氨基葡萄糖基转移酶(GlcNAcT)和半乳糖凝集素-1。先前描述的主要表面糖蛋白,谷胱甘肽过氧化物酶(gp29,GPX-1)和细胞因子同源物巨噬细胞迁移抑制因子(MIF-1)。释放最丰富的蛋白质之一是磷酸丙糖异构酶(TPI),但仅在体提取物中发现了许多其他糖酵解酶(如醛缩酶和GAPDH)。在BES中鉴定出的最杰出的新产品中,有11种小运甲状腺素蛋白样蛋白和3种富含谷氨酰胺的重复粘蛋白样蛋白。值得注意的是,没有证据表明存在任何与马来芽孢杆菌中Wolbachia内共生体的基因组相对应的分泌蛋白。用抗磷酸胆碱(PC)单克隆抗体进行的Western印迹鉴定出Blc中主要的PC携带蛋白是GlcNAcT,而不是ES-62同源物,而用人丝虫病血清进行的检测显示出对galectin-1和加工形式的优先反应性肌动蛋白。总的来说,该分析表明选择性释放了一组新鉴定的蛋白质,这些蛋白质以前不怀疑与宿主-寄生虫的界面有关,并为丝虫的生物学提供了重要的新观点。

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