National Institutes of Health/National Institute of General Medical Sciences
RO1 GM111506
United States
Citation
Journal: Nat Methods / Year: 2020 Title: In situ structure determination at nanometer resolution using TYGRESS. Authors: Kangkang Song / Zhiguo Shang / Xiaofeng Fu / Xiaochu Lou / Nikolaus Grigorieff / Daniela Nicastro / Abstract: 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 ...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.
Resolution.type: BY AUTHOR / Resolution: 12.0 Å / Resolution method: FSC 0.143 CUT-OFF / Details: TYGRESS was used for the reconstruction method / Number images used: 18857
Initial angle assignment
Type: OTHER / Details: From subtomogram alignment
Final angle assignment
Type: PROJECTION MATCHING
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