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

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,...

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Detalles Bibliográficos
Autores principales: 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.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8495048/
https://www.ncbi.nlm.nih.gov/pubmed/34642671
http://dx.doi.org/10.1016/j.xpro.2021.100906
Descripción
Sumario: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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