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首页> 外文期刊>Infection and immunity >Surface Immunolabeling and Consensus Computational Framework To Identify Candidate Rare Outer Membrane Proteins of Treponema pallidum
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Surface Immunolabeling and Consensus Computational Framework To Identify Candidate Rare Outer Membrane Proteins of Treponema pallidum

机译：表面免疫标记和共识计算框架，以识别梅毒螺旋体的候选稀有外膜蛋白。

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Treponema pallidum reacts poorly with the antibodies present in rabbit and human syphilitic sera, a property attributed to the paucity of proteins in its outer membrane. To better understand the basis for the syphilis spirochete's “stealth pathogenicity,” we used a dual-label, 3-step amplified assay in which treponemes encapsulated in gel microdroplets were probed with syphilitic sera in parallel with anti-FlaA antibodies. A small (approximately 5 to 10%) but reproducible fraction of intact treponemes bound IgG and/or IgM antibodies. Three lines of evidence supported the notion that the surface antigens were likely β-barrel-forming outer membrane proteins (OMPs): (i) surface labeling with anti-lipoidal (VDRL) antibodies was not observed, (ii) immunoblot analysis confirmed prior results showing that T. pallidum glycolipids are not immunoreactive, and (iii) labeling of intact organisms was not appreciably affected by proteinase K (PK) treatment. With this method, we also demonstrate that TprK (TP0897), an extensively studied candidate OMP, and TP0136, a lipoprotein recently reported to be surface exposed, are both periplasmic. Consistent with the immunolabeling studies, TprK was also found to lack amphiphilicity, a characteristic property of β-barrel-forming proteins. Using a consensus computational framework that combined subcellular localization and β-barrel structural prediction tools, we generated ranked groups of candidate rare OMPs, the predicted T. pallidum outer membrane proteome (OMPeome), which we postulate includes the surface-exposed molecules detected by our enhanced gel microdroplet assay. In addition to underscoring the syphilis spirochete's remarkably poor surface antigenicity, our findings help to explain the complex and shifting balance between pathogen and host defenses that characterizes syphilitic infection.
机译：梅毒螺旋体与兔和人的梅毒血清中的抗体反应不良，这归因于其外膜中蛋白质的缺乏。为了更好地了解梅毒螺旋体“隐身致病性”的基础，我们使用了双标记，三步扩增试验，在该试验中，梅毒血清与抗FlaA抗体平行探测了封装在凝胶微滴中的色氨酸。一小部分（约5％至10％）但完整的可重复组蛋白与IgG和/或IgM抗体结合。有三点证据支持表面抗原可能是形成β桶的外膜蛋白（OMP）的观点：（i）未观察到抗脂质体（VDRL）抗体的表面标记，（ii）免疫印迹分析证实了先前的结果显示该 T。苍白球糖脂没有免疫反应，并且（iii）完整的生物体的标记未受到蛋白酶K（PK）处理的明显影响。通过这种方法，我们还证明了广泛研究的候选OMP TprK（TP0897）和最近报道表面暴露的脂蛋白TP0136都是周质的。与免疫标记研究一致，还发现TprK缺乏两亲性，这是β桶形成蛋白的特征。使用结合亚细胞定位和β桶结构预测工具的共识计算框架，我们生成了候选稀有OMP（预测的 T）排名组。我们假设的苍白球外膜蛋白质组（OMPeome）包括通过增强凝胶微滴测定法检测到的表面暴露分子。除了强调梅毒螺旋体的表面抗原性极差外，我们的发现还有助于解释病原体与宿主防御之间复杂而不断变化的平衡，而这些平衡是梅毒感染的特征。

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