Wang Yaxin, Liu Jingwei, Cao Lin, Wang Wenjing, Sun Yuna, Yin Zheng, Lou Zhiyong
College of Pharmacy and State Key Laboratory of Elemento-Organic Chemistry, Nankai University, Tianjin, 300071, China.
Center of Basic Molecular Science, Department of Chemistry, Tsinghua University, Beijing, 100084, China.
Chembiochem. 2018 Jul 16;19(14):1465-1470. doi: 10.1002/cbic.201800051. Epub 2018 May 18.
Imaging live virus to monitor the viral entry process is essential to understand virus-host interactions during pathogen infection. However, methods for efficient labeling of live viruses, in particular labeling non-enveloped viruses and tracing virus entry processes, remain limited. Recently, labeling by using organometallic palladium reagents has provided a highly efficient and selective way to bioconjugate cysteines of virus proteins. Here, site-specific bioorthogonal labeling mediated by an organometallic palladium reagent on the surface of live enterovirus-71 (EV71) was used to visualize its entry into live cells. In contrast to currently used immunofluorescence and membrane-anchored dyes, this site-specific and quantitative labeling of live EV71 allows temporal imaging of its entry into host cell membranes on the timescale of seconds with little negative impact on its virulence. This method revealed details of EV71 virus entry and has broad applicability for monitoring virus entry that is difficult to assess by using conventional protein-labeling approaches.
对活病毒进行成像以监测病毒进入过程对于理解病原体感染期间的病毒-宿主相互作用至关重要。然而，有效标记活病毒的方法，特别是标记无包膜病毒和追踪病毒进入过程的方法仍然有限。最近，使用有机金属钯试剂进行标记提供了一种高效且选择性的方法来生物共轭病毒蛋白的半胱氨酸。在此，利用有机金属钯试剂介导的在活肠道病毒71型（EV71）表面的位点特异性生物正交标记来可视化其进入活细胞的过程。与目前使用的免疫荧光和膜锚定染料相比，这种对活EV71的位点特异性和定量标记允许在秒级时间尺度上对其进入宿主细胞膜的过程进行实时成像，且对其毒力几乎没有负面影响。该方法揭示了EV71病毒进入的细节，并且对于监测难以通过传统蛋白质标记方法评估的病毒进入具有广泛的适用性。
