Sofia University "St. Kliment Ohridski", Faculty of Physics, 5 James Boucher Blvd., 1164, Sofia, Bulgaria.
Department of Virology, National Centre of Infectious and Parasitic Diseases, 44A Gen. Stoletov Blvd., 1233, Sofia, Bulgaria.
Anal Biochem. 2023 Jun 1;670:115137. doi: 10.1016/j.ab.2023.115137. Epub 2023 Mar 29.
Chemiluminescence was used to test the susceptibility of the SARS-CoV-2 N and S proteins to oxidation by reactive oxygen species (ROS) at pH 7.4 and pH 8.5. The Fenton's system generates various ROS (HO, OH, OH, OOH). All proteins were found to significantly suppress oxidation (the viral proteins exhibited 25-60% effect compared to albumin). In the second system, HO was used both as a strong oxidant and as a ROS. A similar effect was observed (30-70%); N protein approached the effect of albumin at physiological pH (∼45%). In the O-generation system, albumin was most effective in the suppression of generated radicals (75%, pH 7.4). The viral proteins were more susceptible to oxidation (inhibition effect no more than 20%, compared to albumin). The standard antioxidant assay confirmed the strong antioxidant capacity of both viral proteins (1.5-1.7 fold higher than albumin). These results demonstrate the effective and significant inhibition of ROS-induced oxidation by the proteins. Obviously, the viral proteins could not be involved in the oxidative stress reactions during the course of the infection. They even suppress the metabolites involved in its progression. These results can be explained by their structure. Probably, an evolutionary self-defense mechanism of the virus has been developed.
化学发光被用于测试 SARS-CoV-2 的 N 和 S 蛋白在 pH 值为 7.4 和 8.5 时对活性氧(ROS)氧化的敏感性。Fenton 体系会产生各种 ROS(HO、OH、OH、OOH)。所有蛋白都被发现能显著抑制氧化(与白蛋白相比,病毒蛋白的抑制作用为 25-60%)。在第二个体系中,HO 既是强氧化剂,也是 ROS。观察到类似的效果(30-70%);N 蛋白在生理 pH 值(约 45%)下接近白蛋白的效果。在 O 生成体系中,白蛋白在抑制生成的自由基方面最为有效(75%,pH 值为 7.4)。病毒蛋白更容易被氧化(与白蛋白相比,抑制作用不超过 20%)。标准抗氧化测定证实了两种病毒蛋白的强大抗氧化能力(比白蛋白高 1.5-1.7 倍)。这些结果表明蛋白能有效且显著抑制 ROS 诱导的氧化。显然,病毒蛋白不能参与感染过程中的氧化应激反应。它们甚至抑制了与其进展相关的代谢物。这些结果可以用它们的结构来解释。可能病毒已经发展出了一种进化的自我防御机制。