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针对 SARS-CoV-2 核衣壳蛋白的特异性生化和血清学特征分析。

Domain-specific biochemical and serological characterization of SARS-CoV-2 nucleocapsid protein.

机构信息

Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA.

Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63110, USA.

出版信息

STAR Protoc. 2021 Dec 17;2(4):100906. doi: 10.1016/j.xpro.2021.100906. Epub 2021 Oct 7.

DOI:10.1016/j.xpro.2021.100906
PMID:34642671
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8495048/
Abstract

Nucleocapsid proteins are essential for SARS-CoV-2 life cycle. Here, we describe protocols to gather domain-specific insights about essential properties of nucleocapsids. These assays include dynamic light scattering to characterize oligomerization, fluorescence polarization to quantify RNA binding, hydrogen-deuterium exchange mass spectrometry to map RNA binding regions, negative-stain electron microscopy to visualize oligomeric species, interferon reporter assay to evaluate interferon signaling modulation, and a serology assay to reveal insights for improved sensitivity and specificity. These assays are broadly applicable to RNA-encapsidated nucleocapsids. For complete details on the use and execution of this protocol, please refer to Wu et al. (2021).

摘要

核衣壳蛋白是 SARS-CoV-2 生命周期所必需的。在这里，我们描述了一些方案，可用于收集有关核衣壳基本特性的特定领域的见解。这些测定包括动态光散射来表征寡聚化，荧光偏振来定量 RNA 结合，氢氘交换质谱来绘制 RNA 结合区域，负染色电子显微镜来可视化寡聚体种类，干扰素报告基因测定来评估干扰素信号转导的调节，以及血清学测定来揭示提高敏感性和特异性的见解。这些测定广泛适用于 RNA 包裹的核衣壳。有关该方案使用和实施的完整详细信息，请参阅 Wu 等人。（2021 年）。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/adec126a0237/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/06af4f48e889/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/543d4bb06270/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/70b9755ef19a/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/df3c8730dae4/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/1a622cc27820/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/007342e0c378/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/4f89949f1e91/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/0fb2581fe5f3/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/adec126a0237/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/06af4f48e889/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/543d4bb06270/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/70b9755ef19a/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/df3c8730dae4/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/1a622cc27820/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/007342e0c378/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/4f89949f1e91/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/0fb2581fe5f3/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6c/8551226/adec126a0237/gr8.jpg

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本文引用的文献

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Characterization of SARS-CoV-2 nucleocapsid protein reveals multiple functional consequences of the C-terminal domain.
严重急性呼吸综合征冠状病毒2核衣壳蛋白的特征揭示了C末端结构域的多种功能后果。
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