Kominkova Marketa, Michalek Petr, Cihalova Kristyna, Guran Roman, Cernei Natalia, Nejdl Lukas, Smerkova Kristyna, Dostalova Simona, Chudobova Dagmar, Heger Zbynek, Vesely Radek, Gumulec Jaromir, Kynicky Jindrich, Xhaxhiu Kledi, Zitka Ondrej, Adam Vojtech, Kizek Rene
Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno, Czech Republic.
Central European Institute of Technology, Brno University of Technology, Technicka 3058/10, CZ-616 00 Brno, Czech Republic.
Int J Mol Sci. 2015 Mar 31;16(4):7210-29. doi: 10.3390/ijms16047210.
In this work, we focused on the differences between bacterial cultures of E. coli obtained from swabs of infectious wounds of patients compared to laboratory E. coli. In addition, blocking of the protein responsible for the synthesis of glutathione (γ-glutamylcysteine synthase-GCL) using 10 mM buthionine sulfoximine was investigated. Each E. coli showed significant differences in resistance to antibiotics. According to the determined resistance, E. coli were divided into experimental groups based on a statistical evaluation of their properties as more resistant and more sensitive. These groups were also used for finding the differences in a dependence of the glutathione pathway on resistance to antibiotics. More sensitive E. coli showed the same kinetics of glutathione synthesis while blocking GCL (Km 0.1 µM), as compared to non-blocking. In addition, the most frequent mutations in genes of glutathione synthetase, glutathione peroxidase and glutathione reductase were observed in this group compared to laboratory E.coli. The group of "more resistant" E. coli exhibited differences in Km between 0.3 and 0.8 µM. The number of mutations compared to the laboratory E. coli was substantially lower compared to the other group.
在这项研究中,我们重点关注了从患者感染伤口拭子中获取的大肠杆菌培养物与实验室大肠杆菌之间的差异。此外,还研究了使用10 mM丁硫氨酸亚砜胺阻断负责谷胱甘肽合成的蛋白质(γ-谷氨酰半胱氨酸合成酶-GCL)的情况。每种大肠杆菌在抗生素抗性方面都表现出显著差异。根据测定的抗性,基于对其作为抗性更强和敏感性更强的特性的统计评估,将大肠杆菌分为实验组。这些组还用于发现谷胱甘肽途径对抗生素抗性的依赖性差异。与未阻断时相比,更敏感的大肠杆菌在阻断GCL(Km 0.1 µM)时显示出相同的谷胱甘肽合成动力学。此外,与实验室大肠杆菌相比,该组中谷胱甘肽合成酶、谷胱甘肽过氧化物酶和谷胱甘肽还原酶基因中最常见的突变。“抗性更强”的大肠杆菌组在Km值上表现出0.3至0.8 µM的差异。与实验室大肠杆菌相比,该组的突变数量与另一组相比显著更低。