In Situ Cryo-Electron Tomography: A Post-Reductionist Approach to Structural Biology

被引：131

作者：

Asano, Shoh ^{[1
]}

Engel, Benjamin D. ^{[1
]}

Baumeister, Wolfgang ^{[1
]}

机构：

[1] Max Planck Inst Biochem, Dept Mol Struct Biol, Klopferspitz 18, D-82152 Martinsried, Germany

来源：

JOURNAL OF MOLECULAR BIOLOGY | 2016年 / 428卷 / 02期

关键词：

cryo-EM; tomography; template matching; visual proteomics; subtomogram averaging; 3D ELECTRON-MICROSCOPY; CRYO-EM; SAMPLE PREPARATION; VITREOUS SECTIONS; RADIATION-DAMAGE; 26S PROTEASOME; INTACT-CELLS; MOLECULAR ARCHITECTURE; EUKARYOTIC CELLS; LIGHT-MICROSCOPY;

D O I：

10.1016/j.jmb.2015.09.030

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

070307 [化学生物学]; 071010 [生物化学与分子生物学];

摘要：

Cryo-electron tomography is a powerful technique that can faithfully image the native cellular environment at nanometer resolution. Unlike many other imaging approaches, cryo-electron tomography provides a label-free method of detecting biological structures, relying on the intrinsic contrast of frozen cellular material for direct identification of macromolecules. Recent advances in sample preparation, detector technology, and phase plate imaging have enabled the structural characterization of protein complexes within intact cells. Here, we review these technical developments and outline a detailed computational workflow for in situ structural analysis. Two recent studies are described to illustrate how this workflow can be adapted to examine both known and unknown cellular complexes. The stage is now set to realize the promise of visual proteomics a complete structural description of the cell's native molecular landscape. (C) 2015 Elsevier Ltd. All rights reserved.

引用

页码：332 / 343

页数：12

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