Cryo-EM structure of haemoglobin at 3.2 Å determined with the Volta phase plate.
Cryo-EM structure of haemoglobin at 3.2 Å determined with the Volta phase plate.
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DOI：
10.1038/ncomms16099
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发表时间：
2017-06-30
影响因子：
16.6
通讯作者：
Danev R
中科院分区：
文献类型：
--
作者：
Khoshouei M;Radjainia M;Baumeister W;Danev R
随着直接电子探测器的出现，冷冻电子显微镜 (cryo-EM) 的前景发生了深刻的变化。这些相机在处理嵌入非晶冰中的辐射敏感有机材料的固有低对比度和光束引起的运动方面优于以前的探测器，因此它们能够以原子或近原子分辨率对许多大分子组装体进行结构测定。尽管如此，仍然存在局限性，其中之一是目标结构的大小。在这里，我们报告了使用 Volta 相位板来确定 3.2 Å 的人类血红蛋白 (64 kDa) 的结构。我们的结果表明，该方法可以应用于比之前通过传统的基于散焦的方法研究的复合物要小的多的复合物。冷冻电镜现已接近成为晶体学的快速且经济高效的替代方案，用于高分辨率蛋白质结构测定。单颗粒冷冻电镜通常用于大型复合物的结构测定。在这里，作者展示了人类血红蛋白的 3.2 Å 分辨率冷冻电镜结构，其分子量为 64 kDa。
With the advent of direct electron detectors, the perspectives of cryo-electron microscopy (cryo-EM) have changed in a profound way. These cameras are superior to previous detectors in coping with the intrinsically low contrast and beam-induced motion of radiation-sensitive organic materials embedded in amorphous ice, and hence they have enabled the structure determination of many macromolecular assemblies to atomic or near-atomic resolution. Nevertheless, there are still limitations and one of them is the size of the target structure. Here, we report the use of a Volta phase plate in determining the structure of human haemoglobin (64 kDa) at 3.2 Å. Our results demonstrate that this method can be applied to complexes that are significantly smaller than those previously studied by conventional defocus-based approaches. Cryo-EM is now close to becoming a fast and cost-effective alternative to crystallography for high-resolution protein structure determination. Single particle cryo-EM is commonly used for the structure determination of large complexes. Here, the authors present the 3.2 Å resolution cryo-EM structure of human haemoglobin, which has a molecular weight of 64 kDa.