Milestones in electron crystallography |
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Authors: | Ludovic Renault Hui -Ting Chou Po -Lin Chiu Rena M Hill Xiangyan Zeng Bryant Gipson Zi Yan Zhang Anchi Cheng Vinzenz Unger Henning Stahlberg |
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Institution: | (1) Molecular and Cellular Biology, College of Biological Sciences, University of California at Davis, Briggs Hall, 1 Shields Ave., Davis, CA 95616, USA;(2) Department of Cell Biology, The Scripps Research Institute, 1055 N. Torrey Pines Road, La Jolla, CA 02037, USA;(3) Department of Molecular Biophysics and Biochemistry, Yale University, P. O. Box 208024, New Haven, CT 065200-8024, USA |
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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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Keywords: | 2dx Electron crystallography Membrane proteins Image processing 2D crystallization Electron microscopy |
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