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

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

  • STAR Protoc. 2022 May 2;3(2):101406. doi: 10.1016/j.xpro.2022.101406.
Limin Shang 1,  Yuehui Zhang 1,  Yuchen Liu 1,  Chaozhi Jin 1,  Yanan Zhao 1,  Jing Zhang 2,  Pei-Hui Wang 2,  Jian Wang 1
Affiliations

Affiliations

  • 1 State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics, Beijing 102206, China.
  • 2 Key Laboratory for Experimental Teratology of Ministry of Education and Advanced Medical Research Institute, Cheeloo College of Medicine, Shandong University, Jinan, Shandong 250012, China.
PMID: 35611119 DOI: 10.1016/j.xpro.2022.101406
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.

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