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TitleIn situ structure determination at nanometer resolution using TYGRESS.
Journal, issue, pagesNat Methods, Vol. 17, Issue 2, Page 201-208, Year 2020
Publish dateNov 25, 2019
AuthorsKangkang Song / Zhiguo Shang / Xiaofeng Fu / Xiaochu Lou / Nikolaus Grigorieff / Daniela Nicastro /
PubMed AbstractThe resolution of subtomogram averages calculated from cryo-electron tomograms (cryo-ET) of crowded cellular environments is often limited owing to signal loss in, and misalignment of, the ...The resolution of subtomogram averages calculated from cryo-electron tomograms (cryo-ET) of crowded cellular environments is often limited owing to signal loss in, and misalignment of, the subtomograms. By contrast, single-particle cryo-electron microscopy (SP-cryo-EM) routinely reaches near-atomic resolution of isolated complexes. We report a method called 'tomography-guided 3D reconstruction of subcellular structures' (TYGRESS) that is a hybrid of cryo-ET and SP-cryo-EM, and is able to achieve close-to-nanometer resolution of complexes inside crowded cellular environments. TYGRESS combines the advantages of SP-cryo-EM (images with good signal-to-noise ratio and contrast, as well as minimal radiation damage) and subtomogram averaging (three-dimensional alignment of macromolecules in a complex sample). Using TYGRESS, we determined the structure of the intact ciliary axoneme with up to resolution of 12 Å. These results reveal many structural details that were not visible by cryo-ET alone. TYGRESS is generally applicable to cellular complexes that are amenable to subtomogram averaging.
External linksNat Methods / PubMed:31768058 / PubMed Central
MethodsEM (single particle)
Resolution12.0 Å
Structure data

EMDB-9023:
In situ structure of 96-nm repeat unit of Tetrahymena ciliary axoneme by TYGRESS method
Method: EM (single particle) / Resolution: 12.0 Å

Source
  • Tetrahymena thermophila CU428 (eukaryote)

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