Zühlke Daniela, Dörries Kirsten, Bernhardt Jörg, Maaß Sandra, Muntel Jan, Liebscher Volkmar, Pané-Farré Jan, Riedel Katharina, Lalk Michael, Völker Uwe, Engelmann Susanne, Becher Dörte, Fuchs Stephan, Hecker Michael
Institute of Microbiology, Ernst-Moritz-Arndt-University Greifswald, F.-L.-Jahn-Strasse 15, D-17487 Greifswald, Germany.
Institute of Biochemistry, Ernst-Moritz-Arndt-University Greifswald, Felix-Hausdorff-Strasse 4, D-17487 Greifswald, Germany.
Sci Rep. 2016 Jun 27;6:28172. doi: 10.1038/srep28172.
Absolute protein quantification was applied to follow the dynamics of the cytoplasmic proteome of Staphylococcus aureus in response to long-term oxygen starvation. For 1,168 proteins, the majority of all expressed proteins, molecule numbers per cell have been determined to monitor the cellular investments in single branches of bacterial life for the first time. In the presence of glucose the anaerobic protein pattern is characterized by increased amounts of glycolytic and fermentative enzymes such as Eno, GapA1, Ldh1, and PflB. Interestingly, the ferritin-like protein FtnA belongs to the most abundant proteins during anaerobic growth. Depletion of glucose finally leads to an accumulation of different enzymes such as ArcB1, ArcB2, and ArcC2 involved in arginine deiminase pathway. Concentrations of 29 exo- and 78 endometabolites were comparatively assessed and have been integrated to the metabolic networks. Here we provide an almost complete picture on the response to oxygen starvation, from signal transduction pathways to gene expression pattern, from metabolic reorganization after oxygen depletion to beginning cell death and lysis after glucose exhaustion. This experimental approach can be considered as a proof of principle how to combine cell physiology with quantitative proteomics for a new dimension in understanding simple life processes as an entity.
应用绝对蛋白质定量法来追踪金黄色葡萄球菌细胞质蛋白质组在长期缺氧饥饿状态下的动态变化。对于1168种蛋白质（占所有表达蛋白质的大多数），首次确定了每个细胞中的分子数量，以监测细菌生命单个分支中的细胞投入。在有葡萄糖存在的情况下，厌氧蛋白质模式的特征是糖酵解和发酵酶（如烯醇化酶、甘油醛-3-磷酸脱氢酶A1、乳酸脱氢酶1和磷酸转乙酰酶）的含量增加。有趣的是，铁蛋白样蛋白FtnA属于厌氧生长过程中最丰富的蛋白质之一。葡萄糖耗尽最终导致参与精氨酸脱亚胺酶途径的不同酶（如ArcB1、ArcB2和ArcC2）积累。对29种胞外代谢物和78种胞内代谢物的浓度进行了比较评估，并将其整合到代谢网络中。在这里，我们提供了一幅关于对缺氧饥饿反应的几乎完整的图景，从信号转导途径到基因表达模式，从缺氧后的代谢重组到葡萄糖耗尽后的细胞死亡和裂解开始。这种实验方法可被视为一种原理证明，即如何将细胞生理学与定量蛋白质组学相结合，以在理解简单生命过程整体方面开辟新的维度。
