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

Highlights

• Proximity labeling to identify high-confidence SARS-CoV-2 interactors

• Enrichment of biotinylated proteins by TurboID using anti-biotin antibody

• A stringent proteomics workflow to filter non-specific co-purified proteins

• Protocol can be applied to other viral-host protein interaction studies

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).