Endothelial targeting of cowpea mosaic virus (CPMV) via surface vimentin.
Endothelial targeting of cowpea mosaic virus (CPMV) via surface vimentin.
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DOI：
10.1371/journal.ppat.1000417
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发表时间：
2009-05
期刊：
影响因子：
6.7
通讯作者：
Manchester M
中科院分区：
文献类型：
--
作者：
Koudelka KJ;Destito G;Plummer EM;Trauger SA;Siuzdak G;Manchester M
豇豆花叶病毒（CPMV）是小核糖核酸病毒超家族中的一种植物豇豆花叶病毒属病毒，被广泛用于多种生物医学和材料科学应用。尽管其复制仅限于植物，但CPMV能结合并进入哺乳动物细胞，包括内皮细胞，特别是体内的肿瘤新生血管内皮细胞。这种天然特性使得CPMV被用作血管发育活体成像的传感器。CPMV与内皮细胞的结合是通过与一种54kD的细胞表面蛋白相互作用发生的，但这种蛋白此前尚未被鉴定出来。在此我们将CPMV结合蛋白鉴定为中间丝波形蛋白的一种细胞表面形式。通过蛋白质组学筛选确定了CPMV - 波形蛋白的相互作用，并通过CPMV与纯化的波形蛋白直接相互作用以及在波形蛋白敲除细胞系中的抑制作用得到了证实。波形蛋白和CPMV在小鼠和大鼠体内的血管内皮细胞中也共定位。这些研究共同表明，表面波形蛋白介导结合，并可能导致CPMV在体内的内化，确立了表面波形蛋白作为纳米颗粒靶向肿瘤的重要血管内皮配体。这些结果还确立了波形蛋白作为植物界和动物界中小核糖核酸病毒的配体。由于细菌病原体和其他几类病毒也与表面波形蛋白结合，这些研究表明表面波形蛋白在病原体传播中具有共同作用。
豇豆花叶病毒（CPMV）是一种不在动物体内复制的植物病毒，被广泛用于材料科学和纳米生物技术。在口服或静脉注射后，以及在使用荧光修饰形式的CPMV进行的活体血管成像研究中，发现CPMV能与哺乳动物细胞特异性相互作用。已表明CPMV与哺乳动物细胞的结合是通过一种细胞表面结合蛋白（CPMV - BP）。在此我们通过生化分析、活细胞实验和动物模型鉴定了这种细胞表面的CPMV - BP。我们发现这种暴露在表面的蛋白是波形蛋白。波形蛋白主要是一种细胞骨架蛋白，在细胞内部发挥作用，调节结构和动态。我们的结果现在表明表面波形蛋白可作为血管内皮标记物以及这些细胞外表面的靶向选择。这项工作还通过在感染周期中共同使用波形蛋白，统一了CPMV与密切相关的哺乳动物病毒如脊髓灰质炎病毒、泰勒氏鼠脑脊髓炎病毒（TMEV）和柯萨奇病毒之间的关系。其他几种细菌和病毒病原体也使用表面波形蛋白作为附着受体，这项研究可能会促使开发抑制感染的广谱策略。
Cowpea mosaic virus (CPMV) is a plant comovirus in the picornavirus superfamily, and is used for a wide variety of biomedical and material science applications. Although its replication is restricted to plants, CPMV binds to and enters mammalian cells, including endothelial cells and particularly tumor neovascular endothelium in vivo. This natural capacity has lead to the use of CPMV as a sensor for intravital imaging of vascular development. Binding of CPMV to endothelial cells occurs via interaction with a 54 kD cell-surface protein, but this protein has not previously been identified. Here we identify the CPMV binding protein as a cell-surface form of the intermediate filament vimentin. The CPMV-vimentin interaction was established using proteomic screens and confirmed by direct interaction of CPMV with purified vimentin, as well as inhibition in a vimentin-knockout cell line. Vimentin and CPMV were also co-localized in vascular endothelium of mouse and rat in vivo. Together these studies indicate that surface vimentin mediates binding and may lead to internalization of CPMV in vivo, establishing surface vimentin as an important vascular endothelial ligand for nanoparticle targeting to tumors. These results also establish vimentin as a ligand for picornaviruses in both the plant and animal kingdoms of life. Since bacterial pathogens and several other classes of viruses also bind to surface vimentin, these studies suggest a common role for surface vimentin in pathogen transmission. Cowpea mosaic virus (CPMV), a plant virus that does not replicate in animals, is extensively used in material science and nanobiotechnology. CPMV has been found to specifically interact with mammalian cells after oral or intravenous administration, as well as in intravital vascular imaging studies that used a fluorescently modified form of CPMV. Binding of CPMV to mammalian cells was shown to be via a cell-surface binding protein (CPMV-BP). Herein we identify this cell surface CPMV-BP through biochemical analysis, live cell experiments, and animal models. We found this surface exposed protein to be vimentin. Vimentin is principally a cytoskeletal protein that functions in the interior of cells to modulate architecture and dynamics. Our results now indicate surface vimentin can be used as a vascular endothelial marker and targeting option on the exterior surface of these cells. This work also unifies the relationship between CPMV and closely related mammalian viruses such as poliovirus, Theiler's murine encephalomyelitis virus (TMEV), and coxsackie virus through the collective use of vimentin during their infectious cycle. Several other bacterial and viral pathogens use surface vimentin as an attachment receptor as well, and this research may lead to the development of broad-spectrum strategies to inhibit infection.
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