The depressed P cycle contributes to the acquisition of ampicillin resistance in Edwardsiella piscicida
The depressed P cycle contributes to the acquisition of ampicillin resistance in Edwardsiella piscicida
复制标题
P循环抑制导致杀鱼爱德华氏菌获得氨苄西林耐药性
DOI：
10.1016/j.jprot.2019.103562
复制
发表时间：
2020-02-10
影响因子：
3.3
通讯作者：
Li, Hui
中科院分区：
文献类型：
--
作者：
Su, Yu-bin;Kuang, Su-fang;Li, Hui
抗生素耐药细菌对人类健康和水产养殖构成日益严重的威胁。为了进一步探讨细菌抗生素耐药机制，使用 iTRAQ 鉴定了氨苄西林耐药 LTB4 (LTB4-R-AMP)（一种爱德华氏菌）的差异蛋白质组。总共鉴定出 102 个差异蛋白，其中 50 个上调，52 个下调。由于其中许多变化与代谢有关，因此使用交互式途径探索器 (iPath) 来了解 LTB4-R-AMP 中的全局差异代谢反应。该分析将整体代谢调节抑制确定为 LTB4-R-AMP 的最典型特征。 LTB4-R-AMP 中的膜电位和 ATP 低于对照，支持中心碳代谢和能量代谢减少。由于丙酮酸循环（P循环）在中心碳代谢和能量代谢中起着关键作用，进一步研究集中在P循环上，结果表明LTB4-R-AMP中P循环中的基因表达和酶活性降低。这些结果支持以下结论：P 循环抑制有助于杀鱼肠球菌获得氨苄青霉素抗性。这些发现表明，蛋白质组学和 iPath 分析相结合可以提供全局代谢谱，这有助于我们更好地了解氨苄青霉素耐药性与细胞代谢之间的相关性。意义：本研究利用iTRAQ探索Edwardsiella piscicida的氨苄西林耐药机制，发现许多蛋白质丰度差异与代谢有关。 IPath 进一步鉴定了整体抑制的代谢调节，并将丙酮酸循环减少描述为耐氨苄西林杀鱼肠球菌的最典型特征，丙酮酸循环中基因表达和酶活性减少支持了这一点。一致地，检测到较低的膜电位和 ATP。这些结果揭示了氨苄西林耐药的代谢机制，并为通过代谢组学重编程恢复耐药提供了坚实的证据。
Antibiotic-resistant bacteria are an increasingly serious threat to human health and aquaculture. To further explore bacterial antibiotic resistance mechanism, iTRAQ is used to identify a differential proteome in ampicillin-resistant LTB4 (LTB4-R-AMP), a strain of Edwardsiella piscicida. A total of 102 differentially proteins with 50 upregulation and 52 downregulation are identified. Since many of these changes are related to metabolism, interactive pathways explorer(iPath) is used to understand a global differentially metabolic response in LTB4-R-AMP. This analysis identifies a global depressed metabolic modulation as the most characteristic feature of LTB4-R-AMP. Lower membrane potential and ATP in LTB4-R-AMP than control support that the central carbon metabolism and energy metabolism are reduced. Since the pyruvate cycle (the P cycle) plays a key role in the central carbon metabolism and energy metabolism, further investigation focuses on the P cycle and shows that expression of genes and activity of enzymes in the P cycle are decreased in LTB4-R-AMP. These results support the conclusion that the depressed P cycle contributes to the acquisition of ampicillin resistance in E.piscicida. These findings indicate that the combination of proteomics and iPath analysis can provide a global metabolic profile, which helps us better understand the correlation between ampicillin resistance and cellular metabolism. Significance: The present study uses iTRAQ to explore ampicillin resistance mechanism in Edwardsiella piscicida and finds many of these differential abundances of proteins are related to metabolism. IPath further identifies a global depressed metabolic modulation and characterizes the reduced pyruvate cycle as the most characteristic feature of the ampicillin-resistant E. piscicida, which is supported by reduced expression of genes and activity of enzymes in the pyruvate cycle. Consisitently, lower membrane potential and ATP are detetced. These results reveal the metabolic mechanism of ampicillin resistance and provide a solid proof to revert the resistance by reprogramming metabolomics.