Reining in polyoma virus associated nephropathy: design and characterization of a template mimicking BK viral coat protein cellular binding.
Reining in polyoma virus associated nephropathy: design and characterization of a template mimicking BK viral coat protein cellular binding.
复制标题
控制多瘤病毒相关肾病：模拟 BK 病毒外壳蛋白细胞结合的模板的设计和表征。
DOI：
10.1021/bi300639d
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发表时间：
2012
期刊：
影响因子：
2.9
通讯作者：
Mierke,DaleF
中科院分区：
文献类型：
--
作者：
Audu,ChristopherO;O'Hara,Bethany;Pellegrini,Maria;Wang,Lei;Atwood,WalterJ;Mierke,DaleF
BK多瘤病毒是肾移植后慢性排斥反应的主要原因。治疗干预的一个靶点是BK病毒与宿主细胞的初始结合。我们假设BKV感染率可以通过竞争性阻止病毒与细胞结合来抑制。与N-末端糖蛋白复合的同源病毒的X-射线结构表明，病毒外壳的BC和HI环是结合并从而感染宿主细胞的决定因素。病毒外壳的大尺寸使其无法进行常见的生物物理和小分子筛选研究。因此，我们试图开发一种更小的蛋白模板，其以充分模拟BK病毒外壳蛋白与细胞的结合特征的方式掺入所鉴定的BK病毒外壳的结合环。这样的模拟物可以作为用于鉴定BK病毒进展的抑制剂的工具。在此，我们报告的设计和表征的一个减少的大小和可溶性模板来自一个四螺旋的蛋白质的Thermatoga maritimaarchaea细菌的BKV，它保持了拓扑显示的BC和HI环中发现的病毒外壳蛋白，VP 1，BKV。我们证明，GT 1b和GD 1b唾液酸神经节苷脂，结合到BKV的VP 1，也与我们的BKV模板。采用GFP标记的模板，我们显示宿主细胞的关联是剂量依赖性的，可以减少神经氨酸酶治疗。这些数据表明，BKV模板模拟了野生型病毒外壳蛋白VP 1观察到的宿主细胞结合。
The BK polyoma virus is a leading cause of chronic post kidney transplantation rejection. One target for therapeutic intervention is the initial association of the BK virus with the host cell. We hypothesize that the rate of BKV infection can be curbed by competitively preventing viral binding to cells. The X-ray structures of homologous viruses complexed with N-terminal glycoproteins suggest that the BC and HI loops of the viral coat are determinant for binding and thereby infection of the host cell. The large size of the viral coat precludes it from common biophysical and small molecule screening studies. Hence, we sought to develop a smaller protein template incorporating the identified binding loops of the BK viral coat in a manner that adequately mimics the binding characteristics of the BK virus coat protein to cells. Such a mimic may serve as a tool for the identification of inhibitors of BK viral progression. Herein, we report the design and characterization of a reduced-size and soluble template derived from a four-helix proteinTM1526 ofThermatoga maritimaarchaea bacteriawhich maintains the topological display of the BC and HI loops as found in the viral coat protein, VP1, of BKV. We demonstrate that the GT1b and GD1b sialogangliosides, which bind to the VP1 of BKV, also associate with our BKV template. Employing a GFP-tagged template, we show host cell association that is dose dependent and that can be reduced by neuraminidase treatment. These data demonstrate that the BKV template mimics the host cell binding observed for the wild-type virus coat protein VP1.