低温电子显微技术在膜蛋白结构研究中的应用和展望.pdfVIP

第23卷 第11期 生命科学 Vol. 23, No. 11 2011年11月 Chinese Bulletin of Life Sciences Nov., 2011 文章编号：1004-0374(2011) 11-1130-10 ∙ 技术与应用∙ 低温电子显微技术在膜蛋白结构研究中的应用和展望 孙　飞*，王　雪 ( 中国科学院生物物理研究所生物大分子国家重点实验室，北京 100101) 摘　要：介绍了蛋白质电子晶体学和单颗粒分析技术这两种低温电子显微技术在膜蛋白和膜蛋白复合体结 构研究中的具体方法和近 10~20 年来的实际应用，并分别分析了这两种方法的优势和瓶颈。此外，还介绍 了Amphipol 替代、Streptavidin 二维晶体锚定脂质体和纳米球包被脂质体等近两年来出现的新的用于低温 电镜成像的膜蛋白样品制备方法。最后对膜蛋白的低温电子显微研究的未来发展做了展望。 关键词：膜蛋白；低温电子显微技术；电子晶体学；单颗粒分析；电镜样品制备 Q-33; Q503 中图分类号： 文献标志码 ：A Application and aspect of cryo-electron microscopy for the structural study of membrane proteins SUN Fei*, WANG Xue (National Laboratory of Biomacromolecule, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China) Abstract: Two three-dimensional cryo-electron microscopic techniques, electron crystallography and single particle analysis, and their application for the structural study of membrane proteins and membrane complexes in the recent ten to twenty years were introduced and reviewed. The benefits and drawbacks of those two methods on the membrane protein structural study were discussed accordingly. Besides, the recent appeared new sample preparation techniques for cryo-electron microscopic study of membrane proteins, e.g. the amphipol usage, liposome tethered by 2D crystal of streptavidin and solid-based liposome, were also introduced and discussed. Finally, I gave my personal aspect of the future development of cryo-electron microscopy on its potential of solving the 3D structure of membrane proteins. Key words: membrane p