Super-resolution imaging visualizes the eightfold symmetry of gp210 proteins around the nuclear pore complex and resolves the central channel with nanometer resolution
Super-resolution imaging visualizes the eightfold symmetry of gp210 proteins around the nuclear pore complex and resolves the central channel with nanometer resolution
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DOI：
10.1242/jcs.098822
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发表时间：
2012-02-01
影响因子：
4
通讯作者：
Sauer, Markus
中科院分区：
文献类型：
--
作者：
Loeschberger, Anna;van de Linde, Sebastian;Sauer, Markus
细胞核孔复合物（NPC）是细胞中最复杂的分子机器之一，它控制着所有分子进出细胞核的运输。由于它们对基因表达和细胞骨架组织等细胞过程的重要性，在过去几十年中，主要通过电子显微镜对细胞核孔复合物的结构进行了广泛研究。我们利用直接随机光学重建显微镜（dSTORM）进行超分辨率成像，研究了分离的非洲爪蟾卵母细胞核膜中细胞核孔复合物的结构，其横向分辨率约为15纳米。通过生成数百个细胞核孔复合物的累积超分辨图像，我们确定了细胞核孔复合物中心通道的直径为41±7纳米，并证明了膜整合蛋白gp210呈八重径向对称分布。双色dSTORM实验强调了高度对称的细胞核孔复合物是控制色差校正质量以及测试超分辨率成像方法能力和可靠性的理想模型结构。
One of the most complex molecular machines of cells is the nuclear pore complex (NPC), which controls all trafficking of molecules in and out of the nucleus. Because of their importance for cellular processes such as gene expression and cytoskeleton organization, the structure of NPCs has been studied extensively during the last few decades, mainly by electron microscopy. We have used super-resolution imaging by direct stochastic optical reconstruction microscopy (dSTORM) to investigate the structure of NPCs in isolated Xenopus laevis oocyte nuclear envelopes, with a lateral resolution of similar to 15 nm. By generating accumulated super-resolved images of hundreds of NPCs we determined the diameter of the central NPC channel to be 41 +/- 7 nm and demonstrate that the integral membrane protein gp210 is distributed in an eightfold radial symmetry. Two-color dSTORM experiments emphasize the highly symmetric NPCs as ideal model structures to control the quality of corrections to chromatic aberration and to test the capability and reliability of super-resolution imaging methods.