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

Electron crystallography of membrane proteins: two-dimensional crystallization and screening by electron microscopy.
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膜蛋白的电子晶体学:电子显微镜的二维结晶和筛选。

DOI:
10.1016/j.ymeth.2006.07.011
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发表时间:
2007
期刊:
影响因子:
4.8
通讯作者:
I. Schmidt
I. Schmidt
中科院分区:
生物学3区
文献类型:
--
作者:
I. Schmidt

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通过电子晶体学,可以以不断提高的分辨率获得膜蛋白的结构和功能信息。虽然电子显微镜和图像处理方法开发方面的大量工作在数据收集和分辨率方面取得了巨大进步,但结晶的一般准则首先开始出现。然而,二维结晶本身始终是结构生物学中这种强大方法的限制因素。通过透析去除去污剂的二维结晶是最广泛使用的技术。透析方法需要考虑四个主要因素:蛋白质制剂、去污剂、添加的脂质以及任何脂质成分以及缓冲条件。同样重要的是适当和仔细的筛选来识别二维晶体。
Structural and functional information of membrane proteins at ever-increasing resolution is being obtained by electron crystallography. While a large amount of work on the development of methods for electron microscopy and image processing has resulted in tremendous advances in terms of speed of data collection and resolution, general guidelines for crystallization are first starting to emerge. Yet two-dimensional crystallization itself will always remain the limiting factor of this powerful approach in structural biology. Two-dimensional crystallization through detergent removal by dialysis is the most widely used technique. Four main factors need to be considered for the dialysis method: the protein preparation, the detergent, the lipid added as well as any constituent lipid, and the buffer conditions. Equally important is proper and careful screening to identify two-dimensional crystals.
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发表时间: 2002-04-12
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