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

机译:在限定的添加5%氯化钠的基本培养基中孵育时金黄色葡萄球菌的氨基酸和蛋白质组学适应

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摘要

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.
机译:金黄色葡萄球菌是一种多功能细菌,可以在各种盐浓度下生存和生长。这项研究调查了细胞是否可以在不支持细胞复制的最小温育培养基中对5%NaCl的攻击存活下来。细胞在液体培养物中生长,洗涤,然后在仅含有甘氨酸和葡萄糖作为底物的PBS和微量元素的培养基中于37°C孵育90分钟。将对照细胞与处理组进行比较,将处理组与另外的5%NaCl一起孵育。与对照细胞相比,暴露于5%NaCl的细胞吸收了更多的甘氨酸,两组均消耗了99%的葡萄糖。经NaCl处理的细胞与对照组相比,脯氨酸和谷氨酸的细胞质水平显着较高,丙氨酸和蛋氨酸的水平较低(p <0.05)。在对照和处理过的细胞中,两种主要的细胞质氨基酸,天冬氨酸和甘氨酸的水平保持恒定。蛋白质组学分析显示,对照组和治疗组之间有10种蛋白质显示出不同的反应。蛋白质的减少主要与蛋白质的生物合成,致病性和细胞粘附过程有关。由于在基本培养基中孵育期间细胞数量保持恒定,因此得出结论,没有细胞分裂可支持种群增长。结果提供了证据,即暴露于NaCl处理的基本培养基中的细胞经历了原位稳态变化,以适应新的环境条件。有人提出这代表了表型转变,形成了类似于小菌落变体的细胞,具有较低的代谢率和与致病性相关的关键蛋白水平较低。

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