Domain-specific biochemical and serological characterization of SARS-CoV-2 nucleocapsid protein
| Autor: | Austin B. Moyle, Niloufar Kavian, Asmaa Hachim, J. S. Malik Peiris, Abraham J. Qavi, Henry W. Rohrs, Nicole D. Wagner, Joyce Sweeney-Gibbons, Aidan R. Cole, Sophie A. Valkenburg, Michael L. Gross, Gaya K. Amarasinghe, Daisy W. Leung, Christopher F. Basler, Christopher W Farnsworth, Chao Wu |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Science (General)
viruses Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Immunology Biophysics Computational biology Antiviral Agents Microbiology General Biochemistry Genetics and Molecular Biology Serology Q1-390 Interferon Health Sciences Protocol medicine Coronavirus Nucleocapsid Proteins Humans Nucleocapsid Clinical Protocol Reporter gene Microscopy General Immunology and Microbiology SARS-CoV-2 Chemistry General Neuroscience COVID-19 RNA Protein Biochemistry Phosphoproteins Structural biology RNA Viral Interferons Fluorescence anisotropy Protein Binding medicine.drug |
| Zdroj: | STAR Protocols, Vol 2, Iss 4, Pp 100906-(2021) STAR Protocols |
| ISSN: | 2666-1667 |
| Popis: | 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. Graphical Abstract |
| Databáze: | OpenAIRE |
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