Structural organization of Weibel-Palade bodies revealed by cryo-EM of vitrified endothelial cells
Structural organization of Weibel-Palade bodies revealed by cryo-EM of vitrified endothelial cells
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DOI：
10.1073/pnas.0902977106
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发表时间：
2009-10-13
影响因子：
11.1
通讯作者：
Rosenthal, Peter B.
中科院分区：
文献类型：
--
作者：
Berriman, John A.;Li, Sam;Rosenthal, Peter B.
在内皮细胞中，多功能血糖蛋白血管性血友病因子 (VWF) 被储存在称为 Weibel-Palade 小体 (WPB) 的特殊分泌颗粒中，以便快速胞吐释放。在整个玻璃化人脐静脉内皮细胞 (HUVEC) 的薄外围进行电子冷冻显微镜检查，可直接对 WPB 及其与紧密排列的膜细胞器、膜管和细丝的 3D 网络的相互作用进行成像。图像的傅里叶分析和断层扫描重建表明，VWF 在 WPB 中被包装为螺旋。 VWF 小管的螺旋特征用于识别含有 VWF 的细胞器并表征其在低剂量图像中的旁晶顺序。我们建立了 WPB 的 3D 模型，其中单个 VWF 螺旋可以弯曲，但其中被 WPB 膜紧密包裹的 VWF 小管的旁晶堆积与颗粒的棒状形态相关。
In endothelial cells, the multifunctional blood glycoprotein von Willebrand Factor (VWF) is stored for rapid exocytic release in specialized secretory granules called Weibel-Palade bodies (WPBs). Electron cryomicroscopy at the thin periphery of whole, vitrified human umbilical vein endothelial cells (HUVECs) is used to directly image WPBs and their interaction with a 3D network of closely apposed membranous organelles, membrane tubules, and filaments. Fourier analysis of images and tomographic reconstruction show that VWF is packaged as a helix in WPBs. The helical signature of VWF tubules is used to identify VWF-containing organelles and characterize their paracrystalline order in low dose images. We build a 3D model of a WPB in which individual VWF helices can bend, but in which the paracrystalline packing of VWF tubules, closely wrapped by the WPB membrane, is associated with the rod-like morphology of the granules.