Fluctuation of multiple metabolic pathways is required for Escherichia coli in response to chlortetracycline stress
Fluctuation of multiple metabolic pathways is required for Escherichia coli in response to chlortetracycline stress
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大肠杆菌响应金霉素应激需要多种代谢途径的波动
DOI：
10.1039/c3mb70522f
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发表时间：
2014-01-01
影响因子：
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通讯作者：
Peng, Xuanxian
中科院分区：
文献类型：
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作者：
Lin, Xiangmin;Kang, Liqun;Peng, Xuanxian
随着药物的过度使用和误用，细菌抗生素耐药性已成为一个全球性的挑战。耐药性的几种机制已被揭示，但关于细菌对抗生素的全球反应的信息在很大程度上是缺失的。本研究采用等压标记相对定量和绝对定量标记定量蛋白质组学技术，对大肠杆菌K12 BW25113在氯四环素胁迫下的差异蛋白质组学进行了表征。利用液相色谱-基质辅助激光解吸电离质谱法共鉴定出723个蛋白，其中包括10763个多肽，丰度递减的有184个，递增的有147个。最有趣的是，关键的代谢途径，如三羧酸循环、丙酮酸代谢和糖酵解/糖异生急剧波动，而参与翻译过程的核糖体蛋白复合物在氯四环素胁迫下普遍升高，这是一种众所周知的代偿策略，由于氯四环素对核糖体的作用。进一步的抗菌素敏感性试验利用这些差异蛋白基因缺失的基因修饰突变体验证了差异蛋白在代谢途径中的作用。我们的研究表明，代谢途径的下调是全球反应的一部分，并在抗生素耐药性中发挥重要作用。这些结果表明，恢复这些波动的途径可能成为对抗抗生素耐药细菌的新策略。
Bacterial antibiotic resistance has become a worldwide challenge with the overuse and misuse of drugs. Several mechanisms for the resistance are revealed, but information regarding the bacterial global response to antibiotics is largely absent. In this study, we characterized the differential proteome of Escherichia coli K12 BW25113 in response to chlortetracycline stress using isobaric tags for relative and absolute quantitation labeling quantitative proteomics technology. A total of 723 proteins including 10 763 peptides were identified with 184 decreasing and 147 increasing in abundance by liquid chromatography matrix assisted laser desorption ionization mass spectrometry. Most interestingly, crucial metabolic pathways such as the tricarboxylic acid cycle, pyruvate metabolism and glycolysis/gluconeogenesis sharply fluctuated, while the ribosome protein complexes contributing to the translation process were generally elevated in chlortetracycline stress, which is known for a compensative tactic due to the action of chlortetracycline on the ribosome. Further antimicrobial susceptibility assays validated the role of differential proteins in metabolic pathways using genetically modified mutants of gene deletion of these differential proteins. Our study demonstrated that the down-regulation of metabolic pathways was a part of the global response and played an important role in the antibiotics resistance. These results indicate that reverting of these fluctuated pathways may become a novel strategy to combat antibiotic-resistant bacteria.