van Genderen E, Li Y W, Nederlof I, Abrahams J P
Biophysical Structural Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands.
Acta Crystallogr D Struct Biol. 2016 Jan;72(Pt 1):34-9. doi: 10.1107/S205979831502149X. Epub 2016 Jan 1.
When 300 kV cryo-EM images at Scherzer focus are acquired from ∼ 100 nm thick three-dimensional protein nanocrystals using a Falcon 2 direct electron detector, Fourier transformation can reveal the crystalline lattice to surprisingly high resolutions, even though the images themselves seem to be devoid of any contrast. Here, it is reported how this lattice information can be enhanced by means of a wave finder in combination with Wiener-type maximum-likelihood filtering. This procedure paves the way towards full three-dimensional structure determination at high resolution for protein crystals.
当使用Falcon 2直接电子探测器从约100纳米厚的三维蛋白质纳米晶体中获取处于Scherzer聚焦的300 kV冷冻电镜图像时,尽管图像本身似乎没有任何对比度,但傅里叶变换能够以惊人的高分辨率揭示晶格结构。在此,报告了如何通过波峰查找器结合维纳型最大似然滤波来增强这种晶格信息。该方法为蛋白质晶体的高分辨率全三维结构测定铺平了道路。