Native conformation of human von Willebrand protein. Analysis by electron microscopy and quasi-elastic light scattering.
Native conformation of human von Willebrand protein. Analysis by electron microscopy and quasi-elastic light scattering.
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DOI：
10.1016/s0021-9258(17)39509-1
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发表时间：
1985-07
期刊：
影响因子：
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通讯作者：
H. Slayter;Joseph Loscalzo;P. Bockenstedt;R. I. Handin
中科院分区：
文献类型：
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作者：
H. Slayter;Joseph Loscalzo;P. Bockenstedt;R. I. Handin
通过电子显微镜和准弹性光散射技术分析冯维勒布兰德因子 (vWF)，以评估这种异质聚合血浆糖蛋白的大小和形状。电子显微照片表明，天然 vWF 分子是柔性的线性聚合物，轮廓长度范围为 100 至 1300 nm。在其典型配置中，聚合物自身盘绕，最大直径范围为 60 至 200 nm。在偶尔注意到的未卷曲聚合物中可以辨别出单个重复原体亚基，并且测量为 100 nm X 1.5-2.0 nm。准弹性光散射分析证实，溶液中纯化的 vWF 分子的尺寸和形状的测量结果与电子显微镜技术获得的结果相似。此外，通过准弹性光散射评估的平均斯托克斯半径和平均回转半径在很宽的值范围内直接相关，电子显微照片测量的直径和轮廓长度也是如此，这表明聚合物的整体形状不会随着尺寸的增加而改变。这项研究支持了天然 vWF 分子是柔性线性聚合物的概念。此外，这项研究清楚地表明，通过电子显微照片评估的聚合物构型是溶液中聚合物构型的有效表示。所提供的数据还为明确的重复原体亚基提供了第一个证据。
von Willebrand factor (vWF) was analyzed by electron microscopic and quasi-elastic light scattering techniques in order to evaluate the size and shape of this heterogeneous polymeric plasma glycoprotein. Electron micrographs demonstrated that native vWF molecules are flexible, linear polymers, ranging in contour length from 100 to 1300 nm. In their typical configuration, the polymers were coiled upon themselves with maximal diameters ranging from 60 to 200 nm. Individual repeating protomeric subunits were discernible in occasionally noted, uncoiled polymers and measured 100 nm X 1.5-2.0 nm. Quasi-elastic light scattering analysis confirmed that measurements of the size and shape of purified vWF molecules in solution were similar to those obtained with electron microscopic techniques. In addition, the mean Stokes radius and mean radius of gyration assessed by quasielastic light scattering were directly related over a wide range of values, as were the diameter and contour length measured from electron micrographs, suggesting that the overall shape of polymers does not change with increasing size. This study supports the concept that native vWF molecules are flexible, linear polymers. In addition, this study clearly shows that the polymer configuration assessed from electron micrographs is a valid representation of the configuration of the polymer in solution. The data presented also provide the first evidence for a well-defined, repeating protomeric subunit.