An antibody-based proximity labeling protocol to identify biotinylated interactors of SARS-CoV-2

Abstract

Summary: 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). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Microbiology
• Molecular/Chemical Probes
• Protein Biochemistry
• Proteomics
• Mass Spectrometry