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TitleProgramming Conventional Electron Microscopes for Solving Ultrahigh-Resolution Structures of Small and Macro-Molecules.
Journal, issue, pagesAnal Chem, Vol. 91, Issue 17, Page 10996-11003, Year 2019
Publish dateSep 3, 2019
AuthorsHeng Zhou / Feng Luo / Zhipu Luo / Dan Li / Cong Liu / Xueming Li /
PubMed AbstractMicrocrystal 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.
External linksAnal Chem / PubMed:31334636
MethodsEM (electron crystallography)
Resolution0.6 - 0.67 Å
Structure data

0696

6kj1

EMDB-0696, PDB-6kj1:
200kV MicroED structure of FUS (37-42) SYSGYS solved from merged datasets at 0.65 A
Method: EM (electron crystallography) / Resolution: 0.65 Å

0697

6kj2

EMDB-0697, PDB-6kj2:
200kV MicroED structure of FUS (37-42) SYSGYS solved from single crystal at 0.67 A
Method: EM (electron crystallography) / Resolution: 0.67 Å

0698

6kj3

EMDB-0698, PDB-6kj3:
120kV MicroED structure of FUS (37-42) SYSGYS solved from merged datasets at 0.60 A
Method: EM (electron crystallography) / Resolution: 0.6 Å

0699

6kj4

EMDB-0699, PDB-6kj4:
120kV MicroED structure of FUS (37-42) SYSGYS solved from single crystal at 0.65 A
Method: EM (electron crystallography) / Resolution: 0.65 Å

Chemicals

ChemComp-HOH:
WATER / Water

Source
  • homo sapiens (human)
KeywordsRNA BINDING PROTEIN / FUS / MicroED / Ultrahigh resolution

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