Comparative proteome analysis of Helicobacter pylori
Comparative proteome analysis of Helicobacter pylori
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DOI：
10.1046/j.1365-2958.2000.01896.x
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发表时间：
2000-05-01
影响因子：
3.6
通讯作者：
Meyer, TF
中科院分区：
文献类型：
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作者：
Jungblut, PR;Bumann, D;Meyer, TF
幽门螺杆菌是胃炎、溃疡和胃癌的病原体，感染了世界上大约一半的人口。在对两株菌株26695和J99的全基因组测序后，我们已经接近了研究包含大分子遗传信息的功能部分-蛋白质组的艰巨任务。采用分辨率为5000个蛋白点的高分辨率双向电泳技术，分离了3株幽门螺杆菌26695和J99以及动物模型SS1中使用的突出菌株的蛋白。在琼脂平板上培养5天后，从幽门螺杆菌中分离出1800种蛋白质。利用基质辅助激光解吸/电离质谱（MALDI-MS）多肽质量指纹鉴定了152种蛋白质，包括9种已知的毒力因子和28种抗原。被调查的三个菌株只有几个共同的蛋白质斑点。我们观察到，在二维电泳（2-DE）模式中，具有氨基酸交换的蛋白质只产生一个电荷的净变化。27个预测保守的假设开放阅读框（orf）和6个未知的orf被证实表达。细菌的生长条件被证明对某些蛋白质的存在有影响。一项使用人血清的初步免疫印迹研究表明，这种方法是鉴定具有诊断或治疗价值的蛋白质的理想方法。hp . pylori 2-DE图谱及其鉴定的蛋白种类被添加到动态2D-PAGE数据库（http://www.mpiib-berlin.mpg.de/2D-PAGE/）。蛋白质组学在公共领域的基本知识将成为鉴定新的毒力或致病因素以及对幽门螺杆菌具有潜在诊断或治疗价值的抗原的有效工具。
Helicobacter pylori, the causative agent of gastritis, ulcer and stomach carcinoma, infects approximately half of the worlds population. After sequencing the complete genome of two strains, 26695 and J99, we have approached the demanding task of investigating the functional part of the genetic information containing macromolecules, the proteome. The proteins of three strains of H. pylori, 26695 and J99, and a prominent strain used in animal models SS1, were separated by a high-resolution two-dimensional electrophoresis technique with a resolution power of 5000 protein spots. Up to 1800 protein species were separated from H. pylori which had been cultivated for 5 days on agar plates. Using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) peptide mass fingerprinting we have identified 152 proteins, including nine known virulence factors and 28 antigens. The three strains investigated had only a few protein spots in common. We observe that proteins with an amino acid exchange resulting in a net change of only one charge are shifted in the two-dimensional electrophoresis (2-DE) pattern. The expression of 27 predicted conserved hypothetical open reading frames (ORFs) and six unknown ORFs were confirmed. The growth conditions of the bacteria were shown to have an effect on the presence of certain proteins. A preliminary immunoblotting study using human sera revealed that this approach is ideal for identifying proteins of diagnostic or therapeutic value. H. pylori 2-DE patterns with their identified protein species were added to the dynamic 2D-PAGE database (http://www.mpiib-berlin.mpg.de/2D-PAGE/). This basic knowledge of the proteome in the public domain will be an effective instrument for the identification of new virulence or pathogenic factors, and antigens of potentially diagnostic or curative value against H. pylori.