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).
Keywords: Biophysics; Clinical Protocol; Health Sciences; Immunology; Mass Spectrometry; Microbiology; Microscopy; Molecular Biology; Protein Biochemistry; Protein expression and purification; Structural Biology.
【저자키워드】 immunology, mass spectrometry, Microbiology, Molecular biology, structural biology, Biophysics, Microscopy, Health Sciences, clinical protocol, Protein Biochemistry, Protein expression and purification, 【초록키워드】 immunology, mass spectrometry, Microbiology, Molecular biology, protocol, structural biology, serology, interferon, Biophysics, RNA, Protein, electron microscopy, Sensitivity and specificity, clinical, Microscopy, Biology, interferon signaling, oligomerization, reporter, expression, binding, protocols, life cycle, hydrogen, modulation, SARS-CoV-2 life cycle, mass, Nucleocapsid proteins, protein expression, purification, nucleocapsids, deuterium, Complete, regions, negative-stain electron microscopy, evaluate, include, scattering, 【제목키워드】 Protein, serological, biochemical, SARS-CoV-2 nucleocapsid,