Macromolecular architecture in eukaryotic cells visualized by cryoelectron tomography
Macromolecular architecture in eukaryotic cells visualized by cryoelectron tomography
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DOI：
10.1126/science.1076184
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发表时间：
2002-11-08
期刊：
影响因子：
56.9
通讯作者：
Baumeister, W
中科院分区：
文献类型：
--
作者：
Medalia, O;Weber, I;Baumeister, W
玻璃化细胞的电子断层扫描是一种非侵入性的三维成像技术，为探索细胞质的超分子组织开辟了新的前景。我们将这种技术应用于Dictyosteoblastoma细胞，专注于肌动蛋白细胞骨架。在没有预先去除膜或提取可溶性蛋白质的情况下重建的肌动蛋白网络中，可以分析单个微片段的交联、它们的分支角度和膜附着位点。在5至6纳米的分辨率下，可以在不受干扰的细胞环境中识别具有不同形状的单个大分子，例如26 S蛋白酶体。
Electron tomography of vitrifed cells is a noninvasive three-dimensional imaging technique that opens up new vistas for exploring the supramolecular organization of the cytoplasm. We applied this technique to Dictyostelium cells, focusing on the actin cytoskeleton. In actin networks reconstructed without prior removal of membranes or extraction of soluble proteins, the cross-linking of individual micro laments, their branching angles, and membrane attachment sites can be analyzed. At a resolution of 5 to 6 nanometers, single macromolecules with distinct shapes, such as the 26S proteasome, can be identified in an unperturbed cellular environment.