生物素化
冠状病毒
计算生物学
生物
严重急性呼吸综合征冠状病毒2型(SARS-CoV-2)
生物素
鉴定(生物学)
2019年冠状病毒病(COVID-19)
中东呼吸综合征冠状病毒
细胞生物学
化学
生物化学
医学
传染病(医学专业)
病理
疾病
植物
作者
Philip V’kovski,Silvio Steiner,Volker Thiel
出处
期刊:Methods in molecular biology
日期:2020-01-01
卷期号:: 187-204
被引量:4
标识
DOI:10.1007/978-1-0716-0900-2_14
摘要
Biotin-based proximity labeling circumvents major pitfalls of classical biochemical approaches to identify protein–protein interactions. It consists of enzyme-catalyzed biotin tags ubiquitously apposed on proteins located in close proximity of the labeling enzyme, followed by affinity purification and identification of biotinylated proteins by mass spectrometry. Here we outline the methods by which the molecular microenvironment of the coronavirus replicase/transcriptase complex (RTC), i.e., proteins located within a close perimeter of the RTC, can be determined by different proximity labeling approaches using BirAR118G (BioID), TurboID, and APEX2. These factors represent a molecular signature of coronavirus RTCs and likely contribute to the viral life cycle, thereby constituting attractive targets for the development of antiviral intervention strategies.
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