SARS-CoV-2 核衣壳蛋白的生化特性分析。
Biochemical characterization of SARS-CoV-2 nucleocapsid protein.
机构信息
Department of Obstetrics and Gynecology, The First Affiliated Hospital of USTC, Division of Molecular Medicine, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230001, China; Hefei National Laboratory for Physical Sciences at Microscale, Laboratory of Structural Immunology, CAS Key Laboratory of Innate Immunity and Chronic Disease, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230027, China.
National Center for Protein Science Shanghai, Shanghai Advanced Research Institute, Chinese Academy of Sciences, Shanghai, 201210, China.
出版信息
Biochem Biophys Res Commun. 2020 Jun 30;527(3):618-623. doi: 10.1016/j.bbrc.2020.04.136. Epub 2020 Apr 30.
The nucleocapsid (N) protein is an important antigen for coronavirus, which participate in RNA package and virus particle release. In this study, we expressed the N protein of SARS-CoV-2 and characterized its biochemical properties. Static light scattering, size exclusive chromatography, and small-angle X-ray scattering (SAXS) showed that the purified N protein is largely a dimer in solution. CD spectra showed that it has a high percentage of disordered region at room temperature while it was best structured at 55 °C, suggesting its structural dynamics. Fluorescence polarization assay showed it has non-specific nucleic acid binding capability, which raised a concern in using it as a diagnostic marker. Immunoblot assays confirmed the presence of IgA, IgM and IgG antibodies against N antigen in COVID-19 infection patients' sera, proving the importance of this antigen in host immunity and diagnostics.
核衣壳 (N) 蛋白是冠状病毒的重要抗原，参与 RNA 包装和病毒粒子释放。在这项研究中，我们表达了 SARS-CoV-2 的 N 蛋白，并对其生化特性进行了表征。静态光散射、尺寸排阻色谱和小角 X 射线散射 (SAXS) 表明，纯化的 N 蛋白在溶液中主要是二聚体。圆二色性 (CD) 光谱表明，它在室温下具有很高比例的无规卷曲区域，而在 55°C 时结构最好，表明其结构动态性。荧光偏振测定表明它具有非特异性核酸结合能力，这引起了人们对将其用作诊断标志物的关注。免疫印迹分析证实了 COVID-19 感染患者血清中针对 N 抗原的 IgA、IgM 和 IgG 抗体的存在，证明了该抗原在宿主免疫和诊断中的重要性。
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