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

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

STAR protocols (2021-10-14)
Chao Wu, Abraham J Qavi, Austin B Moyle, Nicole D Wagner, Asmaa Hachim, Niloufar Kavian, Aidan R Cole, Joyce Sweeney-Gibbons, Henry W Rohrs, J S Malik Peiris, Christopher F Basler, Michael L Gross, Sophie A Valkenburg, Christopher W Farnsworth, Gaya K Amarasinghe, Daisy W Leung
PMID34642671
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).

MATERIALS
Product Number
Brand
Product Description
P4417
Sigma-Aldrich
Phosphate buffered saline, tablet
71085
Sigma-Aldrich
KOD DNA Polymerase, High fidelity DNA polymerase designed for accurate PCR amplification of DNA templates for general cloning and cDNA amplification applications.