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TitleProtein and Organic-Molecular Crystallography With 300kV Electrons on a Direct Electron Detector.
Journal, issue, pagesFront Mol Biosci, Vol. 7, Page 612226, Year 2020
Publish dateJan 6, 2021
AuthorsKiyofumi Takaba / Saori Maki-Yonekura / Satoru Inoue / Tatsuo Hasegawa / Koji Yonekura /
PubMed AbstractElectron 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 ...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.
External linksFront Mol Biosci / PubMed:33469549 / PubMed Central
MethodsEM (electron crystallography)
Resolution3.2 Å
Structure data

PDB-7di8:
Electron crystallographic structure of Catalase using a direct electron detector at 300 kV
Method: ELECTRON CRYSTALLOGRAPHY / Resolution: 3.2 Å

Chemicals

ChemComp-HEM:
PROTOPORPHYRIN IX CONTAINING FE / Heme B

ChemComp-NDP:
NADPH DIHYDRO-NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE / Nicotinamide adenine dinucleotide phosphate

Source
  • bos taurus (cattle)
KeywordsOXIDOREDUCTASE / electron 3d crystallography / direct detector / cryo arm / parallem

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