首页> 外文期刊>Acta biomaterialia >Proteomic evaluation of biological nanoparticles isolated from human kidney stones and calcified arteries.
【24h】

Proteomic evaluation of biological nanoparticles isolated from human kidney stones and calcified arteries.

机译:从人肾结石和钙化动脉分离的生物纳米颗粒的蛋白质组学评估。

获取原文
获取原文并翻译 | 示例
           

摘要

Calcifying biological nanoparticles (NPs) develop under cell culture conditions from homogenates of diverse tissue samples displaying extraosseous mineralization, including kidney stones and calcified aneurysms. Probes to definitively identify NPs in biological systems are lacking. Therefore, the aim of this study was to begin to establish a proteomic biosignature of NPs in order to facilitate more definitive investigation of their contribution to disease. Biological NPs derived from human kidney stones and calcified aneurysms were completely decalcified by overnight treatment with ethylenediaminetetraacetic acid or brief incubation in HCl, as evidenced by lack of a calcium shell and of Alizarin Red S staining, by transmission electron microscopy and confocal microscopy, respectively. Decalcified NPs contained numerous proteins, including some from bovine serum and others of prokaryotic origin. Most prominent of the latter group was EF-Tu, which appeared to be identical to EF-Tu from Staphylococcus epidermidis. A monoclonal antibody against human EF-Tu recognized a protein in Western blots of total NP lysate, as well as in intact NPs by immunofluorescence and immunogold EM. Approximately 8% of NPs were quantitatively recognized by the antibody using flow cytometry. Therefore, we have defined methods to reproducibly decalcify biological NPs, and identified key components of their proteome. These elements, including EF-Tu, can be used as biomarkers to further define the processes that mediate propagation of biological NPs and their contribution to disease.
机译:在细胞培养条件下,钙化生物纳米颗粒(NPs)由显示出骨外矿化的各种组织样品的匀浆形成,包括肾结石和钙化的动脉瘤。缺乏用于确定生物系统中NP的探针。因此,本研究的目的是开始建立NP的蛋白质组学生物特征,以便于更明确地调查其对疾病的贡献。通过乙二胺四乙酸过夜处理或在HCl中短暂孵育,可将人肾结石和钙化的动脉瘤的生物NP完全脱钙,这分别由透射电镜和共聚焦显微镜证明缺乏钙壳和茜素红S染色证明。脱钙的NPs包含许多蛋白质,包括一些来自牛血清的蛋白质和其他原核生物的蛋白质。后者中最突出的是EF-Tu,它似乎与表皮葡萄球菌中的EF-Tu相同。抗人EF-Tu的单克隆抗体通过免疫荧光和免疫金EM在总NP裂解物的Western印迹以及完整NP中识别出一种蛋白质。使用流式细胞仪可将抗体定量识别约8%的NP。因此,我们定义了可重现性脱钙生物NP的方法,并确定了其蛋白质组的关键组成部分。这些元素,包括EF-Tu,可以用作生物标志物,以进一步定义介导生物NP传播及其对疾病的贡献的过程。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号