ジャーナル: Anal Chem / 年: 2019 タイトル: Programming Conventional Electron Microscopes for Solving Ultrahigh-Resolution Structures of Small and Macro-Molecules. 著者: Heng Zhou / Feng Luo / Zhipu Luo / Dan Li / Cong Liu / Xueming Li / 要旨: Microcrystal electron diffraction (MicroED) is becoming a powerful tool in determining the crystal structures of biological macromolecules and small organic compounds. However, wide applications of ...Microcrystal electron diffraction (MicroED) is becoming a powerful tool in determining the crystal structures of biological macromolecules and small organic compounds. However, wide applications of this technique are still limited by the special requirement for radiation-tolerated movie-mode camera and the lack of automated data collection methods. Herein, we develop a stage-camera synchronization scheme to minimize the hardware requirements and enable the use of the conventional electron cryo-microscope with a single-frame CCD camera, which ensures not only the acquisition of ultrahigh-resolution diffraction data but also low cost in practice. This method renders the structure determination of both peptide and small organic compounds at ultrahigh resolution up to ∼0.60 Å with unambiguous assignment of nearly all hydrogen atoms. The present work provides a widely applicable solution for routine structure determination of MicroED and demonstrates the capability of the low-end 120 kV microscope with a CCD camera in solving ultrahigh resolution structures of both organic compounds and biological macromolecules.
履歴
登録
2019年7月20日
登録サイト: PDBJ / 処理サイト: PDBJ
改定 1.0
2019年10月2日
Provider: repository / タイプ: Initial release
改定 1.1
2019年11月20日
Group: Data processing / カテゴリ: em_3d_reconstruction / Item: _em_3d_reconstruction.resolution
∠α: 90 ° / ∠β: 90.48 ° / ∠γ: 90 ° / A: 17.15 Å / B: 4.68 Å / C: 17.65 Å / 空間群名: P1211 / 空間群番号: 4
CTF補正
タイプ: NONE
3次元再構成
解像度: 0.65 Å / 解像度の算出法: DIFFRACTION PATTERN/LAYERLINES / 対称性のタイプ: 3D CRYSTAL
原子モデル構築
プロトコル: AB INITIO MODEL / 空間: RECIPROCAL
精密化
解像度: 0.65→8.58 Å / Cor.coef. Fo:Fc: 0.894 / Cor.coef. Fo:Fc free: 0.866 / SU B: 0.657 / SU ML: 0.023 / 交差検証法: THROUGHOUT / ESU R: 0.025 / ESU R Free: 0.027 詳細: HYDROGENS HAVE BEEN USED IF PRESENT IN THE INPUT U VALUES : REFINED INDIVIDUALLY