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

被引:325
作者
Bai, Xiao-chen [1 ]
Fernandez, Israel S. [1 ]
McMullan, Greg [1 ]
Scheres, Sjors H. W. [1 ]
机构
[1] MRC, Mol Biol Lab, Cambridge CB2 2QH, England
来源
ELIFE | 2013年 / 2卷
基金
英国医学研究理事会; 英国惠康基金;
关键词
ELECTRON CRYOMICROSCOPY; RADIATION-DAMAGE; TRANSFER-RNA; MICROSCOPY; ICE; MACROMOLECULES; LIMITATIONS; SPECIMENS; CONTRAST; TILT;
D O I
10.7554/eLife.00461
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
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页数:12
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