Abstract
Elucidating the molecular interactions between virus and host is fundamental to understanding the mechanism of viral pathogenesis. Here, we present a protocol to screen SARS-CoV-2 protein interactors using an antibody-based TurboID proximity labeling approach. This technique directly identifies biotinylated peptides labeled by the TurboID-tagged viral proteins. We describe the steps to prepare biotinylated peptide samples for mass spectrometry analysis and a stringent workflow to identify biotinylated high-confidence interactors of the virus by filtering out non-specific co-purified proteins. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2022).
Keywords: Mass Spectrometry; Microbiology; Molecular/Chemical Probes; Protein Biochemistry; Proteomics.
【저자키워드】 proteomics, mass spectrometry, Microbiology, Protein Biochemistry, Molecular/Chemical Probes, 【초록키워드】 protocol, viral pathogenesis, peptide, Viral proteins, Proteins, virus, mechanism, non-specific, Analysis, mass, molecular interaction, SARS-CoV-2 protein, labeling, Host, Complete, approach, identify, biotinylated, 【제목키워드】 SARS-CoV-2, protocol, labeling, identify, biotinylated,