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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.
Afiliación
  • 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.
Article en En | MEDLINE | ID: mdl-34642671
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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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ARN Viral / Interferones / Nucleocápside / Proteínas de la Nucleocápside de Coronavirus / SARS-CoV-2 / COVID-19 Límite: Humans Idioma: En Revista: STAR Protoc Año: 2021 Tipo del documento: Article País de afiliación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ARN Viral / Interferones / Nucleocápside / Proteínas de la Nucleocápside de Coronavirus / SARS-CoV-2 / COVID-19 Límite: Humans Idioma: En Revista: STAR Protoc Año: 2021 Tipo del documento: Article País de afiliación: Estados Unidos