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

Autor:	Wu C; Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA., Qavi AJ; Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA., Moyle AB; Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63110, USA., Wagner ND; Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63110, USA., Hachim A; HKU-Pasteur Research Pole, School of Public Health, The University of Hong Kong, Hong Kong, China., Kavian N; 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., Cole AR; Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA., Sweeney-Gibbons J; Center for Microbial Pathogenesis, Institute for Biomedical Sciences, Georgia State University, Atlanta, GA 30303, USA., Rohrs HW; Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63110, USA., Peiris JSM; 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., Basler CF; Center for Microbial Pathogenesis, Institute for Biomedical Sciences, Georgia State University, Atlanta, GA 30303, USA., Gross ML; Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63110, USA., Valkenburg SA; HKU-Pasteur Research Pole, School of Public Health, The University of Hong Kong, Hong Kong, China., Farnsworth CW; Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA., Amarasinghe GK; Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA., Leung DW; 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.
Jazyk:	angličtina
Zdroj:	STAR protocols [STAR Protoc] 2021 Dec 17; Vol. 2 (4), pp. 100906. Date of Electronic Publication: 2021 Oct 07.
DOI:	10.1016/j.xpro.2021.100906
Abstrakt:	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). Competing Interests: M.L.G. is an unpaid member of the scientific advisory board of Protein Metrics and GenNext Technologies, two companies commercializing protein footprinting software and tools. (© 2021 The Authors.)
Databáze:	MEDLINE
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