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

STAR Protocols (Dec 2021)

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

Affiliations

Chao Wu: Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA; Corresponding author
Abraham J. Qavi: Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA
Austin B. Moyle: Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63110, USA
Nicole D. Wagner: Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63110, USA
Asmaa Hachim: HKU-Pasteur Research Pole, School of Public Health, The University of Hong Kong, Hong Kong, China
Niloufar Kavian: HKU-Pasteur Research Pole, School of Public Health, The University of Hong Kong, Hong Kong, China; 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; Institut Cochin, INSERM U1016, Université Paris Descartes, Sorbonne Paris Cité, Paris, France
Aidan R. Cole: Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA
Joyce Sweeney-Gibbons: Center for Microbial Pathogenesis, Institute for Biomedical Sciences, Georgia State University, Atlanta, GA 30303, USA
Henry W. Rohrs: Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63110, USA
J.S. Malik Peiris: HKU-Pasteur Research Pole, School of Public Health, The University of Hong Kong, Hong Kong, China; Division of Public Health Laboratory Sciences, School of Public Health, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China
Christopher F. Basler: Center for Microbial Pathogenesis, Institute for Biomedical Sciences, Georgia State University, Atlanta, GA 30303, USA
Michael L. Gross: Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63110, USA
Sophie A. Valkenburg: HKU-Pasteur Research Pole, School of Public Health, The University of Hong Kong, Hong Kong, China; Corresponding author
Christopher W. Farnsworth: Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA; Corresponding author
Gaya K. Amarasinghe: Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA; Corresponding author
Daisy W. Leung: Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA; Department of Internal Medicine, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA; Corresponding author

Journal volume & issue: Vol. 2, no. 4
p. 100906

Abstract

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Summary: 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).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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