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首页> 外文期刊>Journal of bacteriology >Proteomic Analysis and Identification of Streptococcus pyogenes Surface-Associated Proteins
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Proteomic Analysis and Identification of Streptococcus pyogenes Surface-Associated Proteins

机译：化脓性链球菌表面相关蛋白的蛋白质组学分析和鉴定

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Streptococcus pyogenes is a gram-positive human pathogen that causes a wide spectrum of disease, placing a significant burden on public health. Bacterial surface-associated proteins play crucial roles in host-pathogen interactions and pathogenesis and are important targets for the immune system. The identification of these proteins for vaccine development is an important goal of bacterial proteomics. Here we describe a method of proteolytic digestion of surface-exposed proteins to identify surface antigens of S. pyogenes. Peptides generated by trypsin digestion were analyzed by multidimensional tandem mass spectrometry. This approach allowed the identification of 79 proteins on the bacterial surface, including 14 proteins containing cell wall-anchoring motifs, 12 lipoproteins, 9 secreted proteins, 22 membrane-associated proteins, 1 bacteriophage-associated protein, and 21 proteins commonly identified as cytoplasmic. Thirty-three of these proteins have not been previously identified as cell surface associated in S. pyogenes. Several proteins were expressed in Escherichia coli, and the purified proteins were used to generate specific mouse antisera for use in a whole-cell enzyme-linked immunosorbent assay. The immunoreactivity of specific antisera to some of these antigens confirmed their surface localization. The data reported here will provide guidance in the development of a novel vaccine to prevent infections caused by S. pyogenes.
机译：化脓性链球菌是一种革兰氏阳性人类病原体，可引起多种疾病，给公共卫生带来沉重负担。细菌表面相关蛋白在宿主-病原体相互作用和发病机理中起关键作用，并且是免疫系统的重要靶标。鉴定用于疫苗开发的这些蛋白质是细菌蛋白质组学的重要目标。在这里，我们描述了一种蛋白水解消化表面蛋白的方法，以鉴定 S的表面抗原。化脓。通过多维串联质谱分析胰蛋白酶消化产生的肽。这种方法可以鉴定细菌表面上的79种蛋白质，包括14种含有细胞壁锚定基序的蛋白质，12种脂蛋白，9种分泌蛋白质，22种膜相关蛋白质，1种噬菌体相关蛋白质和21种通常被鉴定为细胞质的蛋白质。这些蛋白质中的33种先前尚未鉴定为与 S相关的细胞表面。化脓。几种蛋白在大肠埃希菌中表达，纯化的蛋白用于产生特异性的小鼠抗血清，用于全细胞酶联免疫吸附试验。特异性抗血清对其中某些抗原的免疫反应证实了它们的表面定位。此处报道的数据将为开发新型疫苗预防 S引起的感染提供指导。化脓。

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