一种基于抗体的邻近标记方案，用于鉴定 SARS-CoV-2 的生物素化相互作用蛋白。
An antibody-based proximity labeling protocol to identify biotinylated interactors of SARS-CoV-2.
机构信息
State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics, Beijing 102206, China.
Key Laboratory for Experimental Teratology of Ministry of Education and Advanced Medical Research Institute, Cheeloo College of Medicine, Shandong University, Jinan, Shandong 250012, China.
出版信息
STAR Protoc. 2022 May 2;3(2):101406. doi: 10.1016/j.xpro.2022.101406. eCollection 2022 Jun 17.
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).
阐明病毒和宿主之间的分子相互作用对于理解病毒发病机制至关重要。在这里，我们提出了一种使用基于抗体的 TurboID 邻近标记方法筛选 SARS-CoV-2 蛋白相互作用物的方案。该技术直接鉴定由 TurboID 标记的病毒蛋白标记的生物素化肽。我们描述了用于质谱分析的生物素化肽样品的制备步骤，并描述了通过过滤非特异性共纯化蛋白来鉴定病毒生物素化高可信度相互作用物的严格工作流程。有关此方案的使用和执行的完整详细信息，请参阅 Zhang 等人。(2022 年)。
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