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

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

Research output: Contribution to journalArticlepeer-review

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).

Original languageEnglish
Article number100906
JournalSTAR Protocols
Volume2
Issue number4
DOIs
StatePublished - Dec 17 2021

Keywords

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

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