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首页> 外文期刊>Clinical proteomics. >Proteome analysis of excretory-secretory proteins of Entamoeba histolytica HM1:IMSS via LC–ESI–MS/MS and LC–MALDI–TOF/TOF
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Proteome analysis of excretory-secretory proteins of Entamoeba histolytica HM1:IMSS via LC–ESI–MS/MS and LC–MALDI–TOF/TOF

机译：通过LC–ESI–MS / MS和LC–MALDI–TOF / TOF对溶组织性变形虫阿米巴HM1：IMSS的分泌蛋白进行蛋白质组分析

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Background Excretory-secretory (ES) proteins of E. histolytica are thought to play important roles in the host invasion, metabolism, and defence. Elucidation of the types and functions of E. histolytica ES proteins can further our understanding of the disease pathogenesis. Thus, the aim of this study is to use proteomics approach to better understand the complex ES proteins of the protozoa. Methods E. histolytica ES proteins were prepared by culturing the trophozoites in protein-free medium. The ES proteins were identified using two mass spectrometry tools, namely, LC–ESI–MS/MS and LC–MALDI–TOF/TOF. The identified proteins were then classified according to their biological processes, molecular functions, and cellular components using the Panther classification system (PantherDB). Results A complementary list of 219 proteins was identified; this comprised 201 proteins detected by LC–ESI–MS/MS and 107 proteins by LC–MALDI–TOF/TOF. Of the 219 proteins, 89 were identified by both mass-spectrometry systems, while 112 and 18 proteins were detected exclusively by LC–ESI–MS/MS and LC–MALDI–TOF/TOF respectively. Biological protein functional analysis using PantherDB showed that 27% of the proteins were involved in metabolic processes. Using molecular functional and cellular component analyses, 35% of the proteins were found to be involved in catalytic activity, and 21% were associated with the cell parts. Conclusion This study showed that complementary use of LC–ESI–MS/MS and LC–MALDI–TOF/TOF has improved the identification of ES proteins. The results have increased our understanding of the types of proteins excreted/secreted by the amoeba and provided further evidence of the involvement of ES proteins in intestinal colonisation and evasion of the host immune system, as well as in encystation and excystation of the parasite.
机译：背景溶组织性大肠杆菌的排泄分泌（ES）蛋白被认为在宿主入侵，代谢和防御中起重要作用。对溶组织性大肠杆菌ES蛋白的类型和功能的阐明可以进一步加深我们对疾病发病机理的了解。因此，本研究的目的是使用蛋白质组学方法更好地了解原生动物的复杂ES蛋白。方法：通过在无蛋白培养基中培养滋养体，制备组织溶大肠杆菌。使用两种质谱分析工具鉴定了ES蛋白，即LC–ESI–MS / MS和LC–MALDI–TOF / TOF。然后使用Panther分类系统（PantherDB）根据其生物过程，分子功能和细胞成分对鉴定出的蛋白质进行分类。结果鉴定出219种蛋白质的互补清单。其中包括LC-ESI-MS / MS检测到的201种蛋白质和LC-MALDI-TOF / TOF检测到的107种蛋白质。在219种蛋白质中，两种质谱系统均鉴定出89种蛋白质，而LC-ESI-MS / MS和LC-MALDI-TOF / TOF分别检测到112种和18种蛋白质。使用PantherDB进行的生物蛋白质功能分析表明，有27％的蛋白质参与了代谢过程。使用分子功能和细胞成分分析，发现35％的蛋白质参与催化活性，而21％的蛋白质与细胞部分相关。结论该研究表明，LC-ESI-MS / MS和LC-MALDI-TOF / TOF的互补使用改善了ES蛋白的鉴定。结果增加了我们对变形虫分泌/分泌的蛋白质类型的了解，并提供了ES蛋白参与肠道定植和逃避宿主免疫系统以及寄生虫侵入和分泌的进一步证据。

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