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Milestones in electron crystallography

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Abstract

Electron crystallography determines the structure of membrane embedded proteins in the two-dimensionally crystallized state by cryo-transmission electron microscopy imaging and computer structure reconstruction. Milestones on the path to the structure are high-level expression, purification of functional protein, reconstitution into two-dimensional lipid membrane crystals, high-resolution imaging, and structure determination by computer image processing. Here we review the current state of these methods. We also created an Internet information exchange platform for electron crystallography, where guidelines for imaging and data processing method are maintained. The server (http://2dx.org) provides the electron crystallography community with a central information exchange platform, which is structured in blog and Wiki form, allowing visitors to add comments or discussions. It currently offers a detailed step-by-step introduction to image processing with the MRC software program. The server is also a repository for the 2dx software package, a user-friendly image processing system for 2D membrane protein crystals.

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Abbreviations

2D:

Two-dimensional

3D:

Three-dimensional

cmc:

Critical micellar concentration

cryo-EM:

Cryo-electron microscopy

GUI:

Graphical user interface

NMR:

Nuclear magnetic resonance

TEM:

Transmission electron microscope

XRD:

X-ray diffraction

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Acknowledgements

This work was in part supported by the NSF, grant number MCB-0447860 and by the NIH, grant number U54-GM074929. V. Unger is supported by NIH grants GM66145 and GM071590. We thank Y. Fujiyoshi and J. Fethière for providing material for Fig. 1.

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Authors and Affiliations

  1. Molecular and Cellular Biology, College of Biological Sciences, University of California at Davis, Briggs Hall, 1 Shields Ave., Davis, CA, 95616, USA

    Ludovic Renault, Hui -Ting Chou, Po -Lin Chiu, Rena M. Hill, Xiangyan Zeng, Bryant Gipson, Zi Yan Zhang & Henning Stahlberg

  2. Department of Cell Biology, The Scripps Research Institute, 1055 N. Torrey Pines Road, La Jolla, CA, 02037, USA

    Anchi Cheng

  3. Department of Molecular Biophysics and Biochemistry, Yale University, P. O. Box 208024, New Haven, CT, 065200-8024, USA

    Vinzenz Unger

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  1. Ludovic Renault
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  2. Hui -Ting Chou
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  10. Henning Stahlberg
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Correspondence to Henning Stahlberg.

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Renault, L., Chou, H.T., Chiu, P.L. et al. Milestones in electron crystallography. J Comput Aided Mol Des 20, 519–527 (2006). https://doi.org/10.1007/s10822-006-9075-x

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  • DOI: https://doi.org/10.1007/s10822-006-9075-x

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