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首页> 外文期刊>Open Journal of Applied Sciences >p54nrb, a PSF Protein Partner, Contributes to Meningitic Escherichia coli K1-Mediated Pathogenicities
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p54nrb, a PSF Protein Partner, Contributes to Meningitic Escherichia coli K1-Mediated Pathogenicities

机译：PSF蛋白合作伙伴p54nrb有助于脑膜炎性大肠杆菌K1介导的致病性

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IbeA is an important invasion determinant contributing to Escherichia coli K1 entry into brain microvascular endothelial cells (BMEC) that is a key step in the pathogenesis of E. coli meningitis. Our previous studies have shown that IbeA-induced signaling and E. coli K1 invasion is mediated by two IbeA-binding proteins, vimentin, which is constitutively present in the surface of human BMECs (HBMECs), and PSF, which is inducibly expressed in both mesenchymal (endothelium) and non-mesenchymal (epithelium) cells. However, it is unknown whether p54nrb, a PSF partner protein, could contribute to the pathogenesis of E. coli K1 meningitis. Here, we reported that a 54-kDa protein was identified by copurification with PSF through IbeA-affinity chromatography as an IbeA-binding protein, which is identical to p54nrb. Both p54nrb and PSF are RNA-binding proteins and share significant sequence homology. The specific interaction between IbeA and p54nrb was confirmed by Western blot and ligand overlay assays. Recombinant p54nrb blocked E. coli K1 invasion of human BMEC very effectively. Overexpressed p54nrb as a GFP fusion protein in the transfected 293T cells significantly enhanced E. coli K1 invasion. Furthermore, higher levels of surface p54nrb in the transfected 293T cells were detected by flow cytometry. These results suggest that the IbeA invasion protein of E. coli K1 interacts with p54nrb for bacterial invasion of human BMEC.
机译：IbeA是导致大肠杆菌K1进入脑微血管内皮细胞（BMEC）的重要入侵决定因素，这是大肠杆菌脑膜炎发病机理中的关键步骤。我们以前的研究表明，IbeA诱导的信号转导和大肠杆菌K1入侵是由两种IbeA结合蛋白介导的，波形蛋白主要存在于人BMEC（HBMEC）的表面，而PSF则可以在两者中诱导表达间充质（上皮）和非间充质（上皮）细胞。但是，尚不清楚p54nrb（一种PSF伴侣蛋白）是否可能导致大肠杆菌K1脑膜炎的发病机理。在这里，我们报道了通过IbeA亲和色谱法与PSF共纯化鉴定出的54 kDa蛋白为IbeA结合蛋白，与p54nrb相同。 p54nrb和PSF都是RNA结合蛋白，并具有明显的序列同源性。 Western印迹和配体覆盖分析证实了IbeA与p54nrb之间的特异性相互作用。重组p54nrb非常有效地阻止了大肠杆菌K1对人BMEC的侵袭。在转染的293T细胞中过表达的p54nrb作为GFP融合蛋白显着增强了大肠杆菌K1的入侵。此外，通过流式细胞术检测到转染的293T细胞中较高水平的表面p54nrb。这些结果表明，大肠杆菌K1的IbeA入侵蛋白与p54nrb相互作用，导致细菌入侵人BMEC。

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