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

Wu, Chao; Qavi, Abraham J; Moyle, Austin B; Wagner, Nicole D; Hachim, Asmaa; Kavian, Niloufar; Cole, Aidan R; Sweeney-Gibbons, Joyce; Rohrs, Henry W; Peiris, J S Malik; Basler, Christopher F; Gross, Michael L; Valkenburg, Sophie A; Farnsworth, Christopher W; Amarasinghe, Gaya K; Leung, Daisy W

Wu, Chao; Qavi, Abraham J; Moyle, Austin B; Wagner, Nicole D; Hachim, Asmaa; Kavian, Niloufar; Cole, Aidan R; Sweeney-Gibbons, Joyce; Rohrs, Henry W; Peiris, J S Malik; Basler, Christopher F; Gross, Michael L; Valkenburg, Sophie A; Farnsworth, Christopher W; Amarasinghe, Gaya K; Leung, Daisy W.

Wu C; Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA.
Qavi AJ; Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA.
Moyle AB; Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63110, USA.
Wagner ND; Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63110, USA.
Hachim A; HKU-Pasteur Research Pole, School of Public Health, The University of Hong Kong, Hong Kong, China.
Kavian N; HKU-Pasteur Research Pole, School of Public Health, The University of Hong Kong, Hong Kong, China.
Cole AR; Université Paris Descartes, Sorbonne Paris Cité, Faculté de Médecine, Assistance Publique-Hôpitaux de Paris, Hôpital Universitaire Paris Centre, Centre Hospitalier Universitaire Cochin, Service d'Immunologie Biologique, Paris, France.
Sweeney-Gibbons J; Institut Cochin, INSERM U1016, Université Paris Descartes, Sorbonne Paris Cité, Paris, France.
Rohrs HW; Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA.
Peiris JSM; Center for Microbial Pathogenesis, Institute for Biomedical Sciences, Georgia State University, Atlanta, GA 30303, USA.
Basler CF; Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63110, USA.
Gross ML; HKU-Pasteur Research Pole, School of Public Health, The University of Hong Kong, Hong Kong, China.
Valkenburg SA; Division of Public Health Laboratory Sciences, School of Public Health, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China.
Farnsworth CW; Center for Microbial Pathogenesis, Institute for Biomedical Sciences, Georgia State University, Atlanta, GA 30303, USA.
Amarasinghe GK; Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63110, USA.
Leung DW; HKU-Pasteur Research Pole, School of Public Health, The University of Hong Kong, Hong Kong, China.

STAR Protoc ; 2(4): 100906, 2021 12 17.

文章在英语 | MEDLINE | ID: covidwho-1458864

ABSTRACT

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).

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Biophysics; Clinical Protocol; Health Sciences; Immunology; Mass Spectrometry; Microbiology; Microscopy; Molecular Biology; Protein Biochemistry; Protein expression and purification; Structural Biology

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全文: 可用 采集: 国际数据库 资料库: MEDLINE 主要主题: RNA, Viral / Interferons / Nucleocapsid / Coronavirus Nucleocapsid Proteins / SARS-CoV-2 / COVID-19 研究类型: 诊断研究 / 实验研究 限制: 人类 语言: 英语 期刊: STAR Protoc 年: 2021 类型: 文章 所属国家: J.xpro.2021.100906

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