Egli Martin
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee, USA.
Curr Protoc Nucleic Acid Chem. 2010 Jun;Chapter 7:Unit 7.13. doi: 10.1002/0471142700.nc0713s41.
A detailed understanding of chemical and biological function and the mechanisms underlying the molecular activities ultimately requires atomic-resolution structural data. Diffraction-based techniques such as single-crystal X-ray crystallography, electron microscopy, and neutron diffraction are well established and they have paved the road to the stunning successes of modern-day structural biology. The major advances achieved in the last 20 years in all aspects of structural research, including sample preparation, crystallization, the construction of synchrotron and spallation sources, phasing approaches, and high-speed computing and visualization, now provide specialists and nonspecialists alike with a steady flow of molecular images of unprecedented detail. The present unit combines a general overview of diffraction methods with a detailed description of the process of a single-crystal X-ray structure determination experiment, from chemical synthesis or expression to phasing and refinement, analysis, and quality control. For novices it may serve as a stepping-stone to more in-depth treatises of the individual topics. Readers relying on structural information for interpreting functional data may find it a useful consumer guide.
对化学和生物学功能以及分子活性背后的机制有详尽的了解最终需要原子分辨率的结构数据。基于衍射的技术,如单晶X射线晶体学、电子显微镜和中子衍射,已经成熟,它们为现代结构生物学的惊人成功铺平了道路。过去20年在结构研究的各个方面取得的重大进展,包括样品制备、结晶、同步加速器和散裂源的建设、相位测定方法以及高速计算和可视化,现在为专家和非专家都提供了源源不断的、细节前所未有的分子图像。本单元结合了衍射方法的概述以及对单晶X射线结构测定实验过程的详细描述,从化学合成或表达,到相位测定、精修、分析和质量控制。对于新手来说,它可以作为深入探讨各个主题的垫脚石。依赖结构信息来解释功能数据的读者可能会发现它是一本有用的实用指南。