Carbon sandwich preparation preserves quality of two-dimensional crystals for cryo-electron microscopy.
Carbon sandwich preparation preserves quality of two-dimensional crystals for cryo-electron microscopy.
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DOI：
10.1093/jmicro/dft038
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发表时间：
2013-12
期刊：
影响因子：
--
通讯作者：
Fujiyoshi Y
中科院分区：
文献类型：
--
作者：
Yang F;Abe K;Tani K;Fujiyoshi Y
电子晶体学是确定膜蛋白结构的重要方法。在本文中，我们展示了碳夹层制备对保存结晶样品质量的影响，使用胃H+，K+-ATP酶的二维（2D）晶体及其分析图像的特征示例。与普通的单碳支撑膜制备相比，碳三明治制备显著地提高了平面二维晶体图像的分辨率。由于水蒸发在碳夹样品中受到限制，这种改善可能是由于保留的水对样品周围环境的剧烈变化（如脱水和盐浓度增加）具有强大的保护作用。碳夹层技术的这种保护作用有助于保持固有的、因此是最佳的分析晶体条件。连同其强大的补偿效果的图像偏移，由于光束诱导的标本充电，碳三明治技术是一个强大的方法，用于保存晶体的膜蛋白具有较大的亲水性区域，如H+，K+-ATP酶，从而构成了一个有效的和高质量的方法收集数据的结构分析这些类型的膜蛋白的电子晶体学。
Electron crystallography is an important method for determining the structure of membrane proteins. In this paper, we show the impact of a carbon sandwich preparation on the preservation of crystalline sample quality, using characteristic examples of two-dimensional (2D) crystals from gastric H+,K+-ATPase and their analyzed images. Compared with the ordinary single carbon support film preparation, the carbon sandwich preparation dramatically enhanced the resolution of images from flat sheet 2D crystals. As water evaporation is restricted in the carbon-sandwiched specimen, the improvement could be due to the strong protective effect of the retained water against drastic changes in the environment surrounding the specimen, such as dehydration and increased salt concentrations. This protective effect by the carbon sandwich technique helped to maintain the inherent and therefore best crystal conditions for analysis. Together with its strong compensation effect for the image shift due to beam-induced specimen charging, the carbon sandwich technique is a powerful method for preserving crystals of membrane proteins with larger hydrophilic regions, such as H+,K+-ATPase, and thus constitutes an efficient and high-quality method for collecting data for the structural analysis of these types of membrane proteins by electron crystallography.
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影响因子：
64.8
作者：
Kimura, Y;Vassylyev, DG;Fujiyoshi, Y
通讯作者：
Fujiyoshi, Y
影响因子：
11.4
作者：
Abe, Kazuhiro;Tani, Kazutoshi;Fujiyoshi, Yoshinori
通讯作者：
Fujiyoshi, Yoshinori
影响因子：
3
作者：
Crowther, RA;Henderson, R;Smith, JM
通讯作者：
Smith, JM
影响因子：
5.6
作者：
Hiroaki, Y;Tani, K;Fujiyoshi, Y
通讯作者：
Fujiyoshi, Y
影响因子：
2.2
作者：
FUJIYOSHI, Y;MIZUSAKI, T;HARADA, Y
通讯作者：
HARADA, Y