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Determination of the ribosome structure to a resolution of 2.5 Å by single‐particle cryo‐EM
Author(s) -
Liu Zheng,
GutierrezVargas Cristina,
Wei Jia,
Grassucci Robert A.,
Sun Ming,
Espioel,
MadisonAntenucci Susan,
Tong Liang,
Frank Joachim
Publication year - 2017
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.3068
Subject(s) - cryo electron microscopy , ribosome , structural biology , single particle analysis , resolution (logic) , eukaryotic ribosome , low resolution , particle (ecology) , computational biology , biology , physics , nanotechnology , biophysics , high resolution , computer science , microbiology and biotechnology , rna , biochemistry , materials science , remote sensing , artificial intelligence , ecology , aerosol , meteorology , gene , geology
Abstract With the advance of new instruments and algorithms, and the accumulation of experience over decades, single‐particle cryo‐EM has become a pivotal part of structural biology. Recently, we determined the structure of a eukaryotic ribosome at 2.5 Å for the large subunit. The ribosome was derived from Trypanosoma cruzi , the protozoan pathogen of Chagas disease. The high‐resolution density map allowed us to discern a large number of unprecedented details including rRNA modifications, water molecules, and ions such as Mg 2+ and Zn 2+ . In this paper, we focus on the procedures for data collection, image processing, and modeling, with particular emphasis on factors that contributed to the attainment of high resolution. The methods described here are readily applicable to other macromolecules for high‐resolution reconstruction by single‐particle cryo‐EM.