Yan Qi, Wei Jingyan, Song Junxia, Li Mengna, Guan Xin, Song Jian
College of Pharmaceutical Science, Jilin University, Changchun 130021, China.
Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130000, China.
Antioxidants (Basel). 2023 Sep 14;12(9):1766. doi: 10.3390/antiox12091766.
Important antioxidant enzymes, glutathione peroxidase (GPx) and superoxide dismutase (SOD), are involved in maintaining redox balance. They can protect each other and result in more efficiently removing excessive reactive oxygen species (ROS), protecting cells against injury, and maintaining the normal metabolism of ROS. In this study, GPx (hGPx1) and GPx (hGPx4) genes were integrated into the same open reading frame with SOD active site (SOD3-72P) genes, respectively, and several novel fusion proteins were obtained by using the UTuT6 expression system for the first time. Among them, Se-hGPx1-L-SOD3-72P is the bifunctional fusion protein with the highest GPx activity and the best anti-hydrogen peroxide inactivation ability thus far. The fusion protein exhibits the strongest alkali and high temperature resistance and a greater protective effect against lipoprotein peroxidation damage. and fusion proteins both have good synergistic and antioxidant abilities in HO-induced RBCs and liver damage models. We believe that this research will help with the development of novel bifunctional fusion proteins and the investigation of the synergistic and catalytic mechanisms of GPx and SOD, which are important in creating novel protein therapeutics.
重要的抗氧化酶，谷胱甘肽过氧化物酶（GPx）和超氧化物歧化酶（SOD），参与维持氧化还原平衡。它们可以相互保护，更有效地清除过量的活性氧（ROS），保护细胞免受损伤，并维持ROS的正常代谢。在本研究中，GPx（hGPx1）和GPx（hGPx4）基因分别与SOD活性位点（SOD3-72P）基因整合到同一个开放阅读框中，并首次使用UTuT6表达系统获得了几种新型融合蛋白。其中，Se-hGPx1-L-SOD3-72P是迄今为止具有最高GPx活性和最佳抗过氧化氢失活能力的双功能融合蛋白。该融合蛋白表现出最强的耐碱性和耐高温性，对脂蛋白过氧化损伤具有更大的保护作用。并且融合蛋白在HO诱导的红细胞和肝损伤模型中均具有良好的协同和抗氧化能力。我们相信这项研究将有助于新型双功能融合蛋白的开发以及GPx和SOD协同和催化机制的研究，这对于创造新型蛋白质疗法具有重要意义。
