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全面分析百日咳博德特氏菌表面蛋白质组并鉴定新的免疫原性蛋白。

A comprehensive analysis of Bordetella pertussis surface proteome and identification of new immunogenic proteins.

机构信息

Department of Biological Sciences, Middle East Technical University, 06531 Ankara, Turkey.

出版信息

Vaccine. 2011 Apr 27;29(19):3583-95. doi: 10.1016/j.vaccine.2011.02.086. Epub 2011 Mar 29.

Abstract

Whooping cough, caused by the gram negative pathogen Bordetella pertussis, is a worldwide acute respiratory disease that predominantly involves infants. In the present study, surface proteins of B. pertussis Tohama I and Saadet strains were identified by using 2DE followed by MALDI-TOF-MS/MS analysis and also geLC-MS/MS. With these approaches it was possible to identify 45 and 226 proteins, respectively. When surface proteins of the strains were separated by 2DE and analyzed by Western blotting for their reactivity, a total of 27 immunogenic spots which correspond to 11 different gene products were determined. Glutamine-binding periplasmic protein, leu/ile/val-binding protein, one putative exported protein, and iron-superoxide dismutase (Fe-SOD) were found as immunogenic for the first time in Bordetella. Of a total of 226 proteins identified, 16 were differentially expressed in B. pertussis Saadet and Tohama I strains. Five proteins were expressed only in Saadet (adhesin, chaperone protein DnaJ, fimbrial protein FimX, putative secreted protein Bsp22 and putative universal stress protein), and two (ABC transporter substrate-binding protein and a putative binding protein-dependent transport periplasmic protein) only in Tohama I.

摘要

百日咳是由革兰氏阴性病原体百日咳博德特氏菌引起的一种全球性急性呼吸道疾病,主要影响婴儿。在本研究中,采用 2DE 结合 MALDI-TOF-MS/MS 分析和 geLC-MS/MS 鉴定了博德特氏菌 Tohama I 和 Saadet 株的表面蛋白。通过这些方法,分别鉴定到 45 和 226 种蛋白质。当用 2DE 分离菌株的表面蛋白并通过 Western blot 分析其反应性时,确定了总共 27 个免疫原性斑点,对应于 11 种不同的基因产物。谷氨酰胺结合周质蛋白、亮氨酸/异亮氨酸/缬氨酸结合蛋白、一种假定的分泌蛋白和铁超氧化物歧化酶(Fe-SOD)首次被发现具有免疫原性。在总共鉴定到的 226 种蛋白质中,16 种在博德特氏菌 Saadet 和 Tohama I 菌株中表达不同。有 5 种蛋白仅在 Saadet 中表达(黏附素、伴侣蛋白 DnaJ、菌毛蛋白 FimX、假定分泌蛋白 Bsp22 和假定普遍应激蛋白),有 2 种(ABC 转运体底物结合蛋白和假定的结合蛋白依赖性转运周质蛋白)仅在 Tohama I 中表达。

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