Structural Study of Heterogeneous Biological Samples by Cryoelectron Microscopy and Image Processing.
Structural Study of Heterogeneous Biological Samples by Cryoelectron Microscopy and Image Processing.
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通过冷冻显微镜和图像处理对异质生物样品的结构研究。
DOI：
10.1155/2017/1032432
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发表时间：
2017
影响因子：
--
通讯作者：
Orlova EV
中科院分区：
文献类型：
--
作者：
White HE;Ignatiou A;Clare DK;Orlova EV
在生物体中，生物大分子具有内在的柔韧性，自然地以多种构象存在。现代电子显微镜，特别是在液氮温度下（cryo-EM），能够在接近自然条件和多种构象状态下可视化生物复合物。在过去的十年中，电子技术和软件开发的进步导致了复合物结构变化的揭示，也提高了EM结构的分辨率。目前，基于单粒子分析（SPA）的结构研究提出了几种分离不同构象状态的方法，从而揭示了配合物的功能机制。解决不同状态的任务需要检查大型数据集、复杂的程序和强大的计算能力。一些方法是基于二维图像的分析，而另一些方法是基于三维研究。在这篇综述中，我们描述了目前在结构构象分析中使用的各种技术中实现的基本原理，并提供了这些方法在生物重要复合物结构研究中成功应用的一些例子。
In living organisms, biological macromolecules are intrinsically flexible and naturally exist in multiple conformations. Modern electron microscopy, especially at liquid nitrogen temperatures (cryo-EM), is able to visualise biocomplexes in nearly native conditions and in multiple conformational states. The advances made during the last decade in electronic technology and software development have led to the revelation of structural variations in complexes and also improved the resolution of EM structures. Nowadays, structural studies based on single particle analysis (SPA) suggests several approaches for the separation of different conformational states and therefore disclosure of the mechanisms for functioning of complexes. The task of resolving different states requires the examination of large datasets, sophisticated programs, and significant computing power. Some methods are based on analysis of two-dimensional images, while others are based on three-dimensional studies. In this review, we describe the basic principles implemented in the various techniques that are currently used in the analysis of structural conformations and provide some examples of successful applications of these methods in structural studies of biologically significant complexes.