Determining structures of RNA aptamers and riboswitches by X-ray crystallography.
Determining structures of RNA aptamers and riboswitches by X-ray crystallography.
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DOI：
10.1007/978-1-59745-557-2_9
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发表时间：
2009
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影响因子：
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通讯作者：
Batey, Robert T
中科院分区：
文献类型：
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作者：
Edwards, Andrea L;Garst, Andrew D;Batey, Robert T
结构生物学在充分理解RNA在生物学中的无数作用方面起着核心作用。近年来，RNA纯化和晶体学方法的创新使得越来越多的独特结构可视化，为其在原子水平上的功能提供了新的见解。本文提出了简化了获得适当折叠和活性RNA的均匀样品的过程的一般方案，这些样品在合适的重原子存在下结晶良好。特别重要的是RNA晶体学的方法，包括探索“构造空间”而不是“条件空间”。此外，开发一种高度灵活的实验分相RNA晶体的方法可能为解决这些结构的相对简单的方法打开大门。
Structural biology plays a central role in gaining a full understanding of the myriad roles of RNA in biology. In recent years, innovative approaches in RNA purification and crystallographic methods have lead to the visualization of an increasing number of unique structures, providing new insights into its function at the atomic level. This article presents general protocols which have streamlined the process of obtaining a homogeneous sample of properly folded and active RNA in high concentrations that crystallizes well in the presence of a suitable heavy-atom for phasing. Of particular importance are approaches toward RNA crystallography that include exploring “construct space” as opposed to “condition space”. Moreover, development of a highly flexible method for experimentally phasing RNA crystals may open the door to a relatively simple means of solving these structures.