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

Authors :
Xiao-chen Bai
Israel S Fernandez
Greg McMullan
Sjors HW Scheres
Source :
eLife, Vol 2 (2013)
Publication Year :
2013
Publisher :
eLife Sciences Publications Ltd, 2013.

Abstract

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.

Details

Language :
English
ISSN :
2050084X
Volume :
2
Database :
Directory of Open Access Journals
Journal :
eLife
Publication Type :
Academic Journal
Accession number :
edsdoj.06a6e64d4e9f4c049863467416ca02e9
Document Type :
article
Full Text :
https://doi.org/10.7554/eLife.00461