Outer membrane proteomics of kanamycin-resistant Escherichia coli identified MipA as a novel antibiotic resistance-related protein
Outer membrane proteomics of kanamycin-resistant Escherichia coli identified MipA as a novel antibiotic resistance-related protein
复制标题
耐卡那霉素大肠杆菌的外膜蛋白质组学鉴定 MipA 是一种新型抗生素耐药性相关蛋白
DOI：
10.1093/femsle/fnv074
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发表时间：
2015-06-01
影响因子：
2.1
通讯作者：
Peng, Xuan-xian
中科院分区：
文献类型：
--
作者：
Zhang, Dan-feng;Li, Hui;Peng, Xuan-xian
抗生素耐药细菌对人类健康和食品安全构成巨大威胁，迫切需要了解对抗这些细菌的耐药机制。在当前的研究中，应用比较蛋白质组学方法来鉴定与卡那霉素耐药性相关的大肠杆菌 K-12 外膜 (OM) 蛋白。质谱和蛋白质印迹结果显示，在卡那霉素抗性大肠杆菌 K-12 菌株中，OM 蛋白 TolC、Tsx 和 OstA 上调，而 MipA、OmpA、FadL 和 OmpW 下调。 tolC (Delta tolC-Km) 的基因缺失导致卡那霉素的最低抑菌浓度 (MIC) 降低 2 倍，mipA (Delta mipA-Km) 的基因缺失导致卡那霉素的 MIC 增加 4 倍。转基因菌株的 MIC 变化可以通过基因互补完全恢复。与野生型菌株相比，Delta ompA-Km的存活能力显着提高，Delta tsx-Km的存活能力显着降低。我们进一步评估了 MipA 对其他四种抗生素（包括萘啶酸、链霉素、氯霉素和金霉素）的作用和表达，这表明 MipA 是一种与抗生素耐药性相关的新型 OM 蛋白。
Antibiotic-resistant bacteria are a great threat to human health and food safety and there is an urgent need to understand the mechanisms of resistance for combating these bacteria. In the current study, comparative proteomic methodologies were applied to identify Escherichia coli K-12 outer membrane (OM) proteins related to kanamycin resistance. Mass spectrometry and western blotting results revealed that OM proteins TolC, Tsx and OstA were up-regulated, whereas MipA, OmpA, FadL and OmpW were down-regulated in kanamycin-resistant E. coli K-12 strain. Genetic deletion of tolC (Delta tolC-Km) led to a 2-fold decrease in the minimum inhibitory concentration (MIC) of kanamycin and deletion of mipA (Delta mipA-Km) resulted in a 4-fold increase in the MIC of kanamycin. Changes in the MICs for genetically modified strains could be completely recovered by gene complementation. Compared with the wild-type strain, the survival capability of Delta ompA-Km was significantly increased and that of Delta tsx-Km was significantly decreased. We further evaluated the role and expression of MipA in response to four other antibiotics including nalidixic acid, streptomycin, chloramphenicol and aureomycin, which suggested that MipA was a novel OM protein related to antibiotic resistance.