2.8 Å resolution reconstruction of the Thermoplasma acidophilum 20S proteasome using cryo-electron microscopy

被引：138

作者：

Campbell, Melody G. ^{[1
,2
]}

Veesler, David ^{[1
,2
,3
]}

Cheng, Anchi ^{[1
,2
,4
]}

Potter, Clinton S. ^{[1
,2
,4
]}

Carragher, Bridget ^{[1
,2
,4
]}

机构：

[1] Scripps Res Inst, Natl Resource Automated Mol Microscopy, La Jolla, CA 92037 USA

[2] Scripps Res Inst, Dept Integrat Struct & Computat Biol, La Jolla, CA 92037 USA

[3] Univ Washington, Dept Biochem, Seattle, WA 98195 USA

[4] New York Struct Biol Ctr, New York, NY USA

来源：

ELIFE | 2015年 / 4卷

基金：

美国国家卫生研究院;

关键词：

EM STRUCTURE DETERMINATION; PARTICLE ELECTRON CRYOMICROSCOPY; BEAM-INDUCED MOTION; CRYO-EM; DENSITY MAPS; CRYSTALLOGRAPHY; VALIDATION; DETECTORS; SPECIMEN; COMPLEX;

D O I：

10.7554/eLife.06380

中图分类号：

Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Recent developments in detector hardware and image-processing software have revolutionized single particle cryo-electron microscopy (cryoEM) and led to a wave of near-atomic resolution (typically similar to 3.3 angstrom) reconstructions. Reaching resolutions higher than 3 angstrom is a prerequisite for structure-based drug design and for cryoEM to become widely interesting to pharmaceutical industries. We report here the structure of the 700 kDa Thermoplasma acidophilum 20S proteasome (T20S), determined at 2.8 angstrom resolution by single-particle cryoEM. The quality of the reconstruction enables identifying the rotameric conformation adopted by some amino-acid side chains (rotamers) and resolving ordered water molecules, in agreement with the expectations for crystal structures at similar resolutions. The results described in this manuscript demonstrate that single particle cryoEM is capable of competing with X-ray crystallography for determination of protein structures of suitable quality for rational drug design.

引用

页码：1 / 22

页数：10

共 48 条

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