Metabolomics Reveal Potential Natural Substrates of AcrB in Escherichia coli and Salmonella enterica Serovar Typhimurium.
Metabolomics Reveal Potential Natural Substrates of AcrB in Escherichia coli and Salmonella enterica Serovar Typhimurium.
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DOI：
10.1128/mbio.00109-21
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发表时间：
2021-03-30
期刊：
影响因子：
6.4
通讯作者：
Piddock LJV
中科院分区：
文献类型：
--
作者：
Wang-Kan X;Rodríguez-Blanco G;Southam AD;Winder CL;Dunn WB;Ivens A;Piddock LJV
多重耐药革兰阴性菌对人类健康构成全球性威胁。AcrB外排泵赋予肠道菌目（包括大肠杆菌和沙门氏菌鼠伤寒血清型）固有和进化的耐药性。在对抗抗生素耐药性的斗争中，靶向细菌耐药性机制的药物可用于恢复抗生素的治疗效果。多药耐药外排复合物AcrAB-TolC是肠球菌目中最具临床相关性的外排泵，是药物发现的靶点。泵蛋白AcrB的抑制允许多种抗生素在细胞内积累，有效地恢复其治疗效力。为了促进AcrB外排抑制剂的开发，需要发现泵的天然底物，因为这些底物可以被化学修饰而成为抑制剂。我们使用非靶向代谢组学方法分析了AcrB在大肠杆菌MG 1655和鼠伤寒沙门氏菌SL 1344中的天然底物谱。我们分析了野生型菌株及其各自的AcrB功能丧失突变体（AcrB D408 A）的内代谢组和外代谢组，以确定AcrB的天然底物代谢物。AcrB蛋白与S.菌和大肠大肠杆菌中，我们观察到大多数不同的代谢反应的外泌体的S。菌和大肠coli AcrB D408 A突变体相对于野生型中的突变体，可能表明这两个物种对相同突变的差异代谢适应。此外，我们还发现了可能与沙门氏菌毒力有关的代谢物种类。鼠伤寒沙门氏菌和AcrB共同的潜在天然底物。
Multidrug-resistant Gram-negative bacteria pose a global threat to human health. The AcrB efflux pump confers inherent and evolved drug resistance to Enterobacterales, including Escherichia coli and Salmonella enterica serovar Typhimurium. In the fight against antibiotic resistance, drugs that target resistance mechanisms in bacteria can be used to restore the therapeutic effectiveness of antibiotics. The multidrug resistance efflux complex AcrAB-TolC is the most clinically relevant efflux pump in Enterobacterales and is a target for drug discovery. Inhibition of the pump protein AcrB allows the intracellular accumulation of a wide variety of antibiotics, effectively restoring their therapeutic potency. To facilitate the development of AcrB efflux inhibitors, it is desirable to discover the native substrates of the pump, as these could be chemically modified to become inhibitors. We analyzed the native substrate profile of AcrB in Escherichia coli MG1655 and Salmonella enterica serovar Typhimurium SL1344 using an untargeted metabolomics approach. We analyzed the endo- and exometabolome of the wild-type strain and their respective AcrB loss-of-function mutants (AcrB D408A) to determine the metabolites that are native substrates of AcrB. Although there is 95% homology between the AcrB proteins of S. Typhimurium and E. coli, we observed mostly different metabolic responses in the exometabolomes of the S. Typhimurium and E. coli AcrB D408A mutants relative to those in the wild type, potentially indicating a differential metabolic adaptation to the same mutation in these two species. Additionally, we uncovered metabolite classes that could be involved in virulence of S. Typhimurium and a potential natural substrate of AcrB common to both species.