Exploration of parameters in cryo-EM leading to an improved density map of the E-coli ribosome

被引：27

作者：

LeBarron, Jamie ^{[2
]}

Grassucci, Robert A. ^{[1
]}

Shaikh, Tanvir R. ^{[2
]}

Baxter, William T. ^{[2
]}

Sengupta, Jayati ^{[2
]}

Frank, Joachim ^{[1
,3
]}

机构：

[1] Columbia Univ, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA

[2] Wadsworth Ctr, Albany, NY 12201 USA

[3] Columbia Univ, Dept Biol Sci, New York, NY 10032 USA

来源：

JOURNAL OF STRUCTURAL BIOLOGY | 2008年 / 164卷 / 01期

关键词：

electron microscopy; single-particle reconstruction; ribosome; resolution;

D O I：

10.1016/j.jsb.2008.05.007

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A number of image processing parameters in the 3D reconstruction of a ribosome complex from a cryo-EM data set were varied to test their effects on the final resolution. The parameters examined were pixel size, window size, and mode of Fourier amplitude enhancement at high spatial frequencies. In addition, the strategy of switching from large to small pixel size during angular refinement was explored. The relationship between resolution (in Fourier space) and the number of particles was observed to follow a lin-log dependence, a relationship that appears to hold for other data, as well. By optimizing the above parameters, and using a lin-log extrapolation to the full data set in the estimation of resolution from half-sets, we obtained a 3D map from 131,599 ribosome particles at 6.7 angstrom resolution (FSC = 0.5). (c) 2008 Elsevier Inc. All rights reserved.

引用

页码：24 / 32

页数：9

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