Electron crystallography of membrane proteins: Two-dimensional crystallization and screening by electron microscopy

Methods ◽  
2007 ◽  
Vol 41 (4) ◽  
pp. 417-426 ◽  
Author(s):  
Ingeborg Schmidt-Krey
Keyword(s):  
Electron Microscopy ◽  
Membrane Proteins ◽  
Two Dimensional ◽  
Electron Crystallography

scholarly journals Automated electron microscopy for evaluating two-dimensional crystallization of membrane proteins

2010 ◽  
Vol 171 (1) ◽  
pp. 102-110 ◽  
Author(s):  
Minghui Hu ◽  
Martin Vink ◽  
Changki Kim ◽  
KD Derr ◽  
John Koss ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Membrane Proteins ◽  
Two Dimensional

scholarly journals Two-Dimensional Crystallization Procedure, from Protein Expression to Sample Preparation

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Qie Kuang ◽  
Pasi Purhonen ◽  
Hans Hebert
Keyword(s):  
Membrane Proteins ◽  
General Idea ◽  
General Description ◽  
Two Dimensional ◽  
Structural Studies ◽  
Electron Crystallography ◽  
Electron Microscopic ◽  
Small Proteins ◽  
Crystallization Procedure ◽  
Related Proteins

Membrane proteins play important roles for living cells. Structural studies of membrane proteins provide deeper understanding of their mechanisms and further aid in drug design. As compared to other methods, electron microscopy is uniquely suitable for analysis of a broad range of specimens, from small proteins to large complexes. Of various electron microscopic methods, electron crystallography is particularly well-suited to study membrane proteins which are reconstituted into two-dimensional crystals in lipid environments. In this review, we discuss the steps and parameters for obtaining large and well-ordered two-dimensional crystals. A general description of the principle in each step is provided since this information can also be applied to other biochemical and biophysical methods. The examples are taken from our own studies and published results with related proteins. Our purpose is to give readers a more general idea of electron crystallography and to share our experiences in obtaining suitable crystals for data collection.

Structure of Membrane Proteins Revealed by Electron Crystallography

2000 ◽  
Vol 6 (S2) ◽  
pp. 234-235
Author(s):  
K. Mitsuoka
Keyword(s):  
Membrane Proteins ◽  
Structure Determination ◽  
Two Dimensional ◽  
Asymmetric Unit ◽  
Electron Crystallography ◽  
Aquaporin 1 ◽  
Diffraction Patterns ◽  
2D Crystals

Because membrane proteins are localized in a continuous lipid bilayer in the native environment, the situation of membrane proteins in the two-dimensional (2D) crystals is quite similar to the environment in vivo. Thus, electron crystallography using 2D crystals is one of the suitable techniques for structure determination of membrane proteins at atomic or near-atomic resolution. Here we describe the structures of the two membrane proteins, bacteriorhodopsin and aquaporin-1, which were solved by electron crystallography at 2.5 and 4.0 Å resolution, respectively.Bacteriorhodopsin (bR) is a light-driven proton pump found in Halobacterium salinarium. The atomic model of the protein was first proposed by electron crystallography and we improved the resolution of the structure determination up to 3.0 Å by collecting 366 electron diffraction patterns and 129 images. The resulted map showed not only a bR molecule but also eight surrounding lipids in the asymmetric unit.

scholarly journals Assessing Two-dimensional Crystallization Trials of Small Membrane Proteins for Structural Biology Studies by Electron Crystallography

10.3791/1846 ◽  
2010 ◽  
Author(s):  
Matthew C. Johnson ◽  
Frederik Rudolph ◽  
Tina M. Dreaden ◽  
Gengxiang Zhao ◽  
Bridgette A. Barry ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Structural Biology ◽  
Two Dimensional ◽  
Electron Crystallography

Testing Parameters for Two-Dimensional Crystallization and Electron Crystallography on Two Eukaryotic Membrane Proteins

2008 ◽  
Vol 14 (S2) ◽  
pp. 1292-1293
Author(s):  
G Zhao ◽  
D Müller ◽  
D Stafford ◽  
Y Kanaoka ◽  
KF Austen ◽  
...  
Keyword(s):  
New Mexico ◽  
Membrane Proteins ◽  
Two Dimensional ◽  
Electron Crystallography ◽  
Eukaryotic Membrane Proteins

Extended abstract of a paper presented at Microscopy and Microanalysis 2008 in Albuquerque, New Mexico, USA, August 3 – August 7, 2008

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