Zernike Phase Contrast Cryo-Electron Microscopy and Tomography for Structure Determination at Nanometer and Subnanometer Resolutions
Name
Murata-2010-Zernike Phase Contra.pdf
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1.15 MB
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Author(s)
Murata, Kazuyoshi
Liu, Xiangan
Danev, Radostin
Jakana, Joanita
Schmid, Michael F.
Nagayama, Kuniaki
Chiu, Wah
King, Jonathan Alan
Date Issued
August 2010
Journal
Structure
Publisher
Elsevier
Citation
Murata, Kazuyoshi, Xiangan Liu, Radostin Danev, Joanita Jakana, Michael F. Schmid, Jonathan King, Kuniaki Nagayama, and Wah Chiu. “Zernike Phase Contrast Cryo-Electron Microscopy and Tomography for Structure Determination at Nanometer and Subnanometer Resolutions.” Structure 18, no. 8 (August 2010): 903–912.
Version
Final published version
Abstract
Zernike phase contrast cryo-electron microscopy (ZPC-cryoEM) is an emerging technique that is capable of producing higher image contrast than conventional cryoEM. By combining this technique with advanced image processing methods, we achieved subnanometer resolution for two biological specimens: 2D bacteriorhodopsin crystal and epsilon15 bacteriophage. For an asymmetric reconstruction of epsilon15 bacteriophage, ZPC-cryoEM can reduce the required amount of data by a factor of ~3, compared with conventional cryoEM. The reconstruction was carried out to 13 Å resolution without the need to correct the contrast transfer function. New structural features at the portal vertex of the epsilon15 bacteriophage are revealed in this reconstruction. Using ZPC cryo-electron tomography (ZPC-cryoET), a similar level of data reduction and higher resolution structures of epsilon15 bacteriophage can be obtained relative to conventional cryoET. These results show quantitatively the benefits of ZPC-cryoEM and ZPC-cryoET for structural determinations of macromolecular machines at nanometer and subnanometer resolutions.
MIT Department
Massachusetts Institute of Technology. Department of Biology
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1016/j.str.2010.06.006
