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在添加了 5%氯化钠的限定性基础培养基中培养时,金黄色葡萄球菌的氨基酸和蛋白质组适应现象。

Amino acids and proteomic acclimation of Staphylococcus aureus when incubated in a defined minimal medium supplemented with 5% sodium chloride.

机构信息

Department of Biology, College of Science, University of Hail, Hail, Saudi Arabia.

Metabolic Research Group, Faculty of Science, School of Environmental and Life Sciences, Callaghan, New South Wales, Australia.

出版信息

Microbiologyopen. 2019 Jun;8(6):e00772. doi: 10.1002/mbo3.772. Epub 2019 Feb 9.

Abstract

Staphylococcus aureus is a versatile bacterium that can adapt to survive and grow in a wide range of salt concentrations. This study investigated whether the cells could mount a response to survive a challenge of 5% NaCl in a minimal incubation medium that would not support cell replication. Cells were grown in liquid culture, washed and then incubated for 90 min at 37°C in a medium that contained only glycine and glucose as substrates in PBS plus trace elements. The control cells were compared with a treatment group which was incubated with an additional 5% NaCl. Significantly more glycine was taken up by the cells exposed to 5% NaCl compared with control cells, and both groups consumed 99% of the glucose supplied. The NaCl treated cells had significantly higher cytoplasmic levels of proline and glutamic acid as well as lower levels of alanine and methionine compared with the controls (p < 0.05). The levels of the two major cytoplasmic amino acids, aspartic acid and glycine, remained constant in control and treated cells. Proteomic analyses revealed that 10 proteins showed differential responses between the control and treatment groups. The reductions in proteins were primarily associated with processes of protein biosynthesis, pathogenicity, and cell adhesion. Since cell numbers remained constant during the incubation period in minimal medium, it was concluded that there was no cell division to support population growth. The results provided evidence that the cells in the minimal medium exposed to the NaCl treatment underwent in situ homeostatic changes to adjust to the new environmental conditions. It was proposed that this represented a phenotypic shift to form cells akin to small colony variants, with lower metabolic rates and lower levels of key proteins associated with pathogenicity.

摘要

金黄色葡萄球菌是一种适应性很强的细菌,能够在广泛的盐浓度下生存和生长。本研究旨在探讨细胞是否能够对 5%氯化钠的挑战做出反应,在不支持细胞复制的最小孵育培养基中存活。细胞在液体培养基中生长,洗涤后在仅含有甘氨酸和葡萄糖作为 PBS 中底物的培养基中于 37°C 孵育 90 分钟,外加痕量元素。将对照细胞与另一个用 5%氯化钠孵育的处理组进行比较。与对照细胞相比,暴露于 5%氯化钠的细胞摄取了更多的甘氨酸,两组均消耗了 99%的葡萄糖供应。与对照组相比,经氯化钠处理的细胞的细胞质中脯氨酸和谷氨酸水平显著升高,丙氨酸和蛋氨酸水平显著降低(p<0.05)。两种主要细胞质氨基酸天冬氨酸和甘氨酸的水平在对照和处理组中保持不变。蛋白质组学分析显示,10 种蛋白质在对照组和处理组之间表现出不同的反应。蛋白质的减少主要与蛋白质生物合成、致病性和细胞黏附过程有关。由于在最小培养基中孵育期间细胞数量保持不变,可以得出结论,没有细胞分裂来支持群体生长。研究结果提供了证据表明,暴露于氯化钠处理的最小培养基中的细胞经历了原位体内平衡变化,以适应新的环境条件。据推测,这代表了一种表型转变,形成了类似于小菌落变体的细胞,其代谢率较低,与致病性相关的关键蛋白质水平较低。

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