Ribosome structures to near-atomic resolution from thirty thousand cryo-EM particles

Although electron cryo-microscopy (cryo-EM) single-particle analysis has become an important tool for structural biology of large and flexible macro-molecular assemblies, the technique has not yet reached its full potential. Besides fundamental limits imposed by radiation damage, poor detectors and...

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Bibliographic Details
Published in:eLife 2013-02, Vol.2, p.e00461-e00461
Main Authors: Bai, Xiao-Chen, Fernandez, Israel S, McMullan, Greg, Scheres, Sjors H W
Format: Article
Language:English
Subjects:
Bayesian
Biophysics and Structural Biology
Cameras
Cryoelectron Microscopy - methods
Direct electron detector
Electron Microscopy
Image processing
Radiation
ribosome
Ribosomes - ultrastructure
Sensors
Standard deviation
Symmetry
T. thermophilus
Thermus thermophilus - metabolism
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Summary:Although electron cryo-microscopy (cryo-EM) single-particle analysis has become an important tool for structural biology of large and flexible macro-molecular assemblies, the technique has not yet reached its full potential. Besides fundamental limits imposed by radiation damage, poor detectors and beam-induced sample movement have been shown to degrade attainable resolutions. A new generation of direct electron detectors may ameliorate both effects. Apart from exhibiting improved signal-to-noise performance, these cameras are also fast enough to follow particle movements during electron irradiation. Here, we assess the potentials of this technology for cryo-EM structure determination. Using a newly developed statistical movie processing approach to compensate for beam-induced movement, we show that ribosome reconstructions with unprecedented resolutions may be calculated from almost two orders of magnitude fewer particles than used previously. Therefore, this methodology may expand the scope of high-resolution cryo-EM to a broad range of biological specimens.DOI:http://dx.doi.org/10.7554/eLife.00461.001.
ISSN:2050-084X
2050-084X
DOI:10.7554/eLife.00461