Department of Biology, Long Island University, Brooklyn, New York, USA.
Antimicrob Agents Chemother. 2013 Nov;57(11):5457-61. doi: 10.1128/AAC.00692-13. Epub 2013 Aug 26.
Glutathione is a tripeptide (l-γ-glutamyl-l-cysteinyl-glycine) thiol compound existing in many bacteria and maintains a proper cellular redox state, thus protecting cells against toxic substances such as reactive oxygen species. Polyamines (spermine and spermidine) are low-molecular-weight aliphatic polycations ubiquitously presenting in all living cells and modulate many cellular functions. We previously reported that exogenous polyamines significantly enhanced β-lactam susceptibility of β-lactam-associated multidrug-resistant Acinetobacter baumannii. In this study, three genes differentially associated with the polyamine effects on β-lactam susceptibility were identified by transposon mutagenesis of A. baumannii ATCC 19606. All three genes encoded components of membrane transport systems. Inactivation of one of the genes encoding a putative glutathione transport ATP-binding protein increased the accumulation of intracellular glutathione (∼150 to ∼200%) and significantly decreased the polyamine effects on β-lactam susceptibility in A. baumannii ATCC 19606. When the cells were grown with polyamines, the levels of intracellular glutathione in A. baumannii ATCC 19606 significantly decreased from ∼0.5 to ∼0.2 nmol, while the levels of extracellular glutathione were correspondingly increased. However, the levels of total glutathione (intra- plus extracellular) were unchanged when the cells were grown with or without polyamines. Overall, these results suggest that exogenous polyamines induce glutathione export, resulting in decreased levels of intracellular glutathione, which may produce an improper cellular redox state that is associated with the polyamine-mediated β-lactam susceptibility of A. baumannii. This finding may provide a clue for development of new antimicrobial agents and/or novel strategies to treat multidrug-resistant A. baumannii.
谷胱甘肽是一种三肽(l-γ-谷氨酰-l-半胱氨酰-甘氨酸)硫醇化合物,存在于许多细菌中,维持适当的细胞氧化还原状态,从而保护细胞免受活性氧等有毒物质的侵害。多胺(精胺和亚精胺)是低分子量脂肪族多阳离子,普遍存在于所有活细胞中,并调节许多细胞功能。我们之前报道过,外源性多胺显著增强了与β-内酰胺相关的多药耐药鲍曼不动杆菌对β-内酰胺的敏感性。在这项研究中,通过对鲍曼不动杆菌 ATCC 19606 的转座子诱变,鉴定出与多胺对β-内酰胺敏感性相关的三个差异表达基因。这三个基因都编码膜转运系统的组成部分。一个编码谷胱甘肽转运 ATP 结合蛋白的基因失活,增加了细胞内谷胱甘肽的积累(约 150 到 200%),并显著降低了鲍曼不动杆菌 ATCC 19606 中多胺对β-内酰胺敏感性的影响。当细胞用多胺生长时,鲍曼不动杆菌 ATCC 19606 细胞内谷胱甘肽的水平从约 0.5 下降到约 0.2 nmol,而细胞外谷胱甘肽的水平相应增加。然而,当细胞用或不用多胺生长时,总谷胱甘肽(细胞内加细胞外)的水平没有变化。总的来说,这些结果表明,外源性多胺诱导谷胱甘肽外排,导致细胞内谷胱甘肽水平降低,可能导致细胞氧化还原状态异常,与多胺介导的鲍曼不动杆菌对β-内酰胺的敏感性有关。这一发现可能为开发新的抗菌剂和/或治疗多药耐药鲍曼不动杆菌的新策略提供线索。