Proteome analyses of Staphylococcus aureus in growing and non-growing cells:: A physiological approach
Proteome analyses of Staphylococcus aureus in growing and non-growing cells:: A physiological approach
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DOI：
10.1016/j.ijmm.2005.08.002
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发表时间：
2005-12-01
影响因子：
4.1
通讯作者：
Hecker, M
中科院分区：
文献类型：
--
作者：
Kohler, C;Wolff, S;Hecker, M
金黄色葡萄球菌是一种多种多样的人类病原体，可引起从伤口感染到心内膜炎、骨髓炎和败血症等多种疾病。为了研究这种病原菌，我们试图用两种不同的方法来分析金黄色葡萄球菌的细胞质蛋白质组：二维凝胶分析结合基质辅助激光电离飞行时间质谱仪和无胶体系的多维液相色谱-质谱仪。通过结合这两种分析，我们鉴定了1123种细胞质蛋白质，它们代表了生物体细胞质蛋白质组的三分之二。使用我们的标准2D凝胶设置(等电点4-7)，我们鉴定了473个蛋白质，覆盖了这个蛋白质组窗口预测的细胞质蛋白质组的大约40%。已鉴定的蛋白质属于多种细胞功能，从转录和翻译机制、三羧酸循环(TCC)、糖酵解和发酵途径到核苷酸、脂肪酸和细胞壁成分的生物合成途径。虽然这种基于凝胶的蛋白质组学方法涵盖了预测金黄色葡萄球菌的大部分代谢途径，但通过无凝胶方法鉴定了650个额外的蛋白质，其中包括碱性或疏水蛋白质。在我们的工作中，我们建立了一种主2D凝胶，使我们能够研究在复杂介质中生长的金黄色葡萄球菌细胞中核心碳代谢的调节。我们对指数生长期细胞和静止期细胞的蛋白质模式的比较表明，指数生长期细胞中参与蛋白质合成、转录和糖酵解的酶数量更多。相反，TCC和糖异生的酶在稳定期增加。有了这个全面的蛋白质组图谱，我们就有了一个重要的工具来更好地了解人类病原体金黄色葡萄球菌的细胞生理学。(C)2005年爱思唯尔股份有限公司。版权所有。
Staphylococcus aureus is a versatile human pathogen causing a wide variety of diseases ranging from wound infection to endocarditis, osteomyelitis, and sepsis. In order to investigate this pathogen, we sought to analyze the cytoplasmic proteome of S. aureus COL by using two different approaches: two-dimensional (213) gel analyses combined with matrix-assisted laser ionization-time of flight mass spectrometry and a gel-free system using multidimensional liquid chromatography followed by mass spectrometry. By combining both analyses we identified 1123 cytoplasmic proteins that represent two-thirds of the cytoplasmic proteome of the organism. With our standard 2D gel setup (pI 4-7) we identified 473 proteins that cover about 40% of the cytoplasmic proteome predicted for this proteomic window. The identified proteins belong to a variety of cellular functions ranging from the transcriptional and translational machinery, tricarboxylic acid cycle (TCC), glycolysis, and fermentation pathways to biosynthetic pathways of nucleotides, fatty acids, and cell wall components. While most of the metabolic pathways predicted for S. aureus were covered by this gel-based proteomics 650 additional proteins were identified by the gel-free approach, among them alkaline or hydrophobic proteins. In our work, we established a master 2D gel that enabled us to study the regulation of core carbon metabolism in S. aureus cells grown in a complex medium. Our comparison of the protein pattern of exponentially growing cells with that of stationary-phase cells revealed a higher amount of enzymes involved in protein synthesis, transcription, and glycolysis in exponentially growing cells. In contrast, enzymes of the TCC and gluconeogenesis are increased at the stationary phase. With this comprehensive proteome map we have an essential tool for a better understanding of cell physiology of the human pathogen, S. aureus. (c) 2005 Elsevier GmbH. All rights reserved.