Two-dimensional crystallization of integral membrane proteins for electron crystallography.
Two-dimensional crystallization of integral membrane proteins for electron crystallography.
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用于电子晶体学的完整膜蛋白的二维结晶。
DOI:
10.1007/978-1-60761-762-4_10
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发表时间:
2010
期刊:
影响因子:
--
通讯作者:
Ubarretxena-Belandia,Iban
中科院分区:
文献类型:
--
作者:
Stokes,DavidL;Rice,WilliamJ;Hu,Minghui;Kim,Changki;Ubarretxena-Belandia,Iban
Although membrane proteins make up 30% of the proteome and are a common target for therapeutic drugs, determination of their atomic structure remains a technical challenge. Electron crystallography represents an alternative to the conventional methods of X-ray diffraction and NMR and relies on the formation of two-dimensional crystals. These crystals are produced by reconstituting purified, detergent-solubilized membrane proteins back into the native environment of a lipid bilayer. This chapter reviews methods for producing two-dimensional crystals and for screening them by negative stain electron microscopy. In addition, we show examples of the different morphologies that are commonly obtained and describe basic image analysis procedures that can be used to evaluate their promise for structure determination by cryoelectron microscopy.
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