Bilecik Şeyh Edebali University, Faculty of Science, Department of Molecular Biology and Genetics TR, Bilecik, TR, 11100, Türkiye.
Bilecik Şeyh Edebali University, Faculty of Engineering, Department of Bioengineering, Bilecik, TR, 11100, Türkiye.
J Biol Phys. 2023 Sep;49(3):309-327. doi: 10.1007/s10867-023-09632-4. Epub 2023 Apr 3.
Copper (Cu), one of the heavy metals, plays a vital role in many complex biochemical reactions as a trace element. However, it often becomes toxic when its concentration in the cell exceeds a certain level. Homeostasis of metals in the cell is primarily related to regulating metal transport into and out of the cell. Therefore, it is thought that porin proteins, which have a role in membrane permeability, may also play a role in developing Cu resistance. This study identified the differences between the molecular profiles of wild-type Escherichia coli W3110 and its seven different porin mutants exposed to Cu ions using attenuated total reflectance (ATR)-Fourier transform infrared (FTIR) spectroscopy. The results showed that the absence of porin genes elicits global changes in the structure and composition of membrane lipids and proteins, in both the absence and presence of Cu. The lack of porin genes significantly elevated the amounts of fatty acids and phospholipids. When the alterations in protein secondary structures were compared, the quantity of amide I proteins was diminished by the presence of Cu. However, the amount of amide II proteins increased in porin mutant groups independent of Cu presence or absence. The DNAs are transformed from B- and Z-form to A-form due to porin mutations and the presence of Cu ions. The lack of porin genes increased polysaccharide content independent of Cu presence. This study can help characterize Cu detoxification efficiency and guide for obtaining active living cells to be used in bioremediation.
铜(Cu)是一种重金属,作为微量元素在许多复杂的生化反应中起着至关重要的作用。然而,当细胞内的浓度超过一定水平时,它通常会变得有毒。细胞内金属的动态平衡主要与调节金属进出细胞的运输有关。因此,人们认为在膜通透性中起作用的孔蛋白也可能在铜抗性的发展中发挥作用。本研究使用衰减全反射(ATR)-傅里叶变换红外(FTIR)光谱法,鉴定了暴露于铜离子的野生型大肠杆菌 W3110 及其七种不同孔蛋白突变体的分子谱之间的差异。结果表明,孔蛋白基因的缺失会引起膜脂和蛋白质结构和组成的全局性变化,无论是否存在 Cu。孔蛋白基因的缺失会显著增加脂肪酸和磷脂的含量。当比较蛋白质二级结构的变化时,由于存在 Cu,酰胺 I 蛋白的数量减少。然而,无论是否存在 Cu,孔蛋白突变体组中酰胺 II 蛋白的含量都会增加。由于孔蛋白突变和 Cu 离子的存在,DNA 从 B-型和 Z-型转变为 A-型。孔蛋白基因的缺失会增加多糖的含量,而与 Cu 的存在无关。本研究可以帮助表征铜解毒效率,并指导获得活性活细胞用于生物修复。