Proteomic analysis of exoproteins expressed by enterotoxigenic Staphylococcus aureus strains
Proteomic analysis of exoproteins expressed by enterotoxigenic Staphylococcus aureus strains
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DOI：
10.1002/pmic.200700965
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发表时间：
2008-06-01
期刊：
影响因子：
3.4
通讯作者：
Malorni, Antonio
中科院分区：
文献类型：
--
作者：
Pocsfalvi, Gabriella;Cacace, Giuseppina;Malorni, Antonio
致病菌将多种毒力因子分泌到细胞外液和细胞表面，对宿主细胞的定植和叛乱起着重要作用，从而反映细菌致病性的程度。为了探索分泌蛋白质组部分中表达的毒力因子，使用基于凝胶的自下而上蛋白质组学方法分析了不同的金黄色葡萄球菌菌株。总共鉴定出 119 种不同的蛋白质。使用基于 1-DE 和 2-DE 的蛋白质组学检测肠毒素基因簇 (egc) 阴性和 seb 基因阳性 S. aurcus 美国典型培养物保藏中心 (ATCC) 14458 菌株。对二维图谱肠毒素区域的详细分析证实，除了高表达的葡萄球菌肠毒素 B (SEB) 之外，还存在先前通过基因分型预测的肠毒素样蛋白 SE1K 和 SE1Q (Sergeev 等人, J. Clin. Microbiol. 2004, 42, 2134-2143)。细胞生长的指数末期（7 小时）和稳定期（24 小时）的外蛋白模式显示出该区域的高度相似性。对五种金黄色葡萄球菌菌株的肠毒素区域进行比较分析，包括两种临床分离株（RIMD 31092 和 A900322）、一种具有高度流行的 EC 阳性操纵子的食品来源菌株（AB-8802）和一种非产肠毒素参考菌株（ROS），揭示了不同已知肠毒素和其他毒力因子以及许多核心外蛋白的存在。此外，首次在蛋白质水平上证明了SEIL (RIMD 31092)和SEIP (A900322)的产生。在所应用的实验条件下，没有鉴定出由egc操纵子基因编码的肠毒素。
Pathogenic bacteria excrete a variety of virulence factors into extracellular edium and to the cell surface which have essential roles in the colonization and insurrection of the host cells, and thus reflect the degree of bacterial pathogenicity. For the exploration of virulence factors expressed in the secreted proteome fraction, different Staphylococcus aureus strains were analyzed using gel-based bottom-up proteomic approach. A total of 119 distinct proteins were identified. for the enterotoxin gene cluster (egc) negative and seb gene positive S. aurcus American Type Culture Collection (ATCC) 14458 strain by the use of one- and 2-DE based proteomics. Detailed analysis of enterotoxin region of the 2-D map confirmed, beside the highly expressed staphylococcal enterotoxin B (SEB), the presence of enterotoxin-like proteins SE1K and SE1Q previously predicted by genotyping (Sergeev et al., J. Clin. Microbiol. 2004, 42, 2134-2143). Exoprotein patterns at the late-exponential (7 h) and stationary (24 h) phases of cellular growth show a high-level similarity in this region. Comparative analysis of enterotoxin region of five S. aureus strains including two clinical isolates (RIMD 31092 and A900322), a food derived strain (AB-8802) with highly prevalent egc positive operon and a nonenterotoxigenic reference strain (ROS) revealed the presence of different known enterotoxins and other virulence factors along with a number of core exoproteins. In addition, production of SElL (RIMD 31092) and SEIP (A900322) was demonstrated for the first time at the protein level. Under the experimental conditions applied none of the enterotoxins encoded by the genes of egc operon was identified.