Metabolic Perturbations in a Bacillus subtilis clpP Mutant during Glucose Starvation.
Metabolic Perturbations in a Bacillus subtilis clpP Mutant during Glucose Starvation.
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DOI：
10.3390/metabo7040063
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发表时间：
2017-11-24
期刊：
影响因子：
4.1
通讯作者：
Lalk M
中科院分区：
文献类型：
--
作者：
Schultz D;Schlüter R;Gerth U;Lalk M
蛋白质水解是所有生物体维持蛋白质稳态和适应不断变化的环境条件所必需的。ClpP是枯草芽孢杆菌中的主要蛋白酶，与不同的Clp ATP酶形成复合物。这些复合物在热应激过程中起着至关重要的作用，而且在孢子形成或细胞形态中也起着重要作用。特别是细胞壁、氨基酸和核酸生物合成的酶是葡萄糖饥饿期间蛋白酶ClpP的已知底物。本研究的目的是分析clpP突变对不同生长阶段代谢的影响，并寻找推定的新ClpP底物。因此，B。在化学成分确定的培养基中培养subtilis 168细胞和一个等基因突变株，并通过1H-NMR、HPLC-MS和GC-MS的组合分析代谢物组。此外，通过电子显微镜观察细胞形态。与野生型相比，clpP突变体显示出更高水平的大多数糖酵解代谢产物、柠檬酸循环的中间体、氨基酸和肽聚糖前体。在clpP突变体的外代谢物组中可以检测到溢出代谢物的强烈分泌。此外，观察到磷壁酸代谢产物CDP-甘油的大量增加，并伴有细胞壁肿胀。我们的研究结果表明，代谢组和相应的蛋白质组数据的B之间的可识别的相关性。subtilis clpP突变体。此外，我们的研究结果表明ClpP对负责磷壁酸生物合成的Tag蛋白的影响。
Proteolysis is essential for all living organisms to maintain the protein homeostasis and to adapt to changing environmental conditions. ClpP is the main protease in Bacillus subtilis, and forms complexes with different Clp ATPases. These complexes play crucial roles during heat stress, but also in sporulation or cell morphology. Especially enzymes of cell wall-, amino acid-, and nucleic acid biosynthesis are known substrates of the protease ClpP during glucose starvation. The aim of this study was to analyze the influence of a clpP mutation on the metabolism in different growth phases and to search for putative new ClpP substrates. Therefore, B. subtilis 168 cells and an isogenic ∆clpP mutant were cultivated in a chemical defined medium, and the metabolome was analyzed by a combination of 1H-NMR, HPLC-MS, and GC-MS. Additionally, the cell morphology was investigated by electron microscopy. The clpP mutant showed higher levels of most glycolytic metabolites, the intermediates of the citric acid cycle, amino acids, and peptidoglycan precursors when compared to the wild-type. A strong secretion of overflow metabolites could be detected in the exo-metabolome of the clpP mutant. Furthermore, a massive increase was observed for the teichoic acid metabolite CDP-glycerol in combination with a swelling of the cell wall. Our results show a recognizable correlation between the metabolome and the corresponding proteome data of B. subtilis clpP mutant. Moreover, our results suggest an influence of ClpP on Tag proteins that are responsible for teichoic acids biosynthesis.
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影响因子：
3.7
作者：
Dörries K;Lalk M
通讯作者：
Lalk M
影响因子：
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作者：
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作者：
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