Department of Engineering, Gustav Wieds Vej 10, Aarhus University, 8000, Aarhus C, Denmark.
Department of Molecular biology, Gustav Wieds Vej 10, Aarhus University, 8000, Aarhus C, Denmark.
Sci Rep. 2017 Jun 15;7(1):3576. doi: 10.1038/s41598-017-03799-2.
In recent years, there has been an increasing appreciation of the importance of secreted and extracellular proteins that traditionally have been considered as intracellular components. Vimentin is a highly abundant intermediate filament protein, and its intracellular functions have been investigated in a large number of studies. Recently, however, vimentin has been shown to take part in significant processes outside the cell. Our understanding of the functions of extracellular vimentin is, however, limited. In this study we demonstrate that a vimentin specific antibody, obtained by phage antibody technology, promotes tube formation of endothelial cells in a 2D matrigel assay. By binding vimentin, the antibody increases the tube formation by 21% after 5 hours of incubation. Addition of the antibody directly to cultured endothelial cells does not influence endothelial cell migration or proliferation. The enhanced tube formation can be seen for up to 10 hours where after the effect decreases. It is shown that the antibody-binding site is located on the coil 2 domain of vimentin. To our knowledge this is the first study that demonstrates an enhanced tube formation by binding vimentin in a 2D matrigel assay under normoxic conditions.
近年来,人们越来越意识到传统上被认为是细胞内成分的分泌蛋白和细胞外蛋白的重要性。波形蛋白是一种丰度很高的中间丝蛋白,已有大量研究对其细胞内功能进行了研究。然而,最近发现波形蛋白参与了细胞外的重要过程。然而,我们对细胞外波形蛋白功能的理解是有限的。在这项研究中,我们证明了一种通过噬菌体抗体技术获得的针对波形蛋白的特异性抗体,可促进内皮细胞在二维基质胶测定中的管状形成。通过与波形蛋白结合,该抗体在孵育 5 小时后可使管状形成增加 21%。将抗体直接添加到培养的内皮细胞中不会影响内皮细胞的迁移或增殖。增强的管状形成可长达 10 小时,之后效果降低。结果表明,抗体结合位点位于波形蛋白的卷曲 2 结构域。据我们所知,这是第一项在常氧条件下通过在二维基质胶测定中结合波形蛋白来增强管状形成的研究。