Protein and Organic-Molecular Crystallography With 300kV Electrons on a Direct Electron Detector.
Protein and Organic-Molecular Crystallography With 300kV Electrons on a Direct Electron Detector.
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DOI：
10.3389/fmolb.2020.612226
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发表时间：
2020
影响因子：
5
通讯作者：
Yonekura K
中科院分区：
文献类型：
--
作者：
Takaba K;Maki-Yonekura S;Inoue S;Hasegawa T;Yonekura K
由于电子的强散射能力，电子三维晶体学可以从各种样品的小尺寸晶体中揭示原子结构。在此，使用JEOL公司的ARM 300型电子显微镜，对直接电子检测器DE 64进行蛋白质和有机分子的小而薄的晶体的检测。该显微镜配备了一个冷场发射枪在300千伏的加速电压下运行，四聚光透镜平行照明，一个列内能量过滤器，和一个稳定的旋转测角器阶段。以无人监督的方式从血红素结合酶过氧化氢酶和代表性有机半导体材料Ph-BTBT-C10的晶体中收集旋转衍射数据。过氧化氢酶的分子置换和Ph-BTBT-C10的直接方法确定的结构。分析表明，该系统工作良好，这些分子的电子3D晶体学具有较小的破坏性，较小的点扩散，和更少的噪声比使用传统的耦合相机。
Electron 3D crystallography can reveal the atomic structure from undersized crystals of various samples owing to the strong scattering power of electrons. Here, a direct electron detector DE64 was tested for small and thin crystals of protein and an organic molecule using a JEOL CRYO ARM 300 electron microscope. The microscope is equipped with a cold-field emission gun operated at an accelerating voltage of 300 kV, quad condenser lenses for parallel illumination, an in-column energy filter, and a stable rotational goniometer stage. Rotational diffraction data were collected in an unsupervised manner from crystals of a heme-binding enzyme catalase and a representative organic semiconductor material Ph-BTBT-C10. The structures were determined by molecular replacement for catalase and by the direct method for Ph-BTBT-C10. The analyses demonstrate that the system works well for electron 3D crystallography of these molecules with less damaging, a smaller point spread, and less noise than using the conventional scintillator-coupled camera.