Determining biomolecular structures near room temperature using X-ray crystallography: concepts, methods and future optimization.
Determining biomolecular structures near room temperature using X-ray crystallography: concepts, methods and future optimization.
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DOI：
10.1107/s2059798322011652
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发表时间：
2023-01-01
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综述和讨论了在室温下以及在∼2 0 0和∼35 0 K之间的温度下从生物分子系统中收集结晶学数据的关键物理原理和方法。在大约二十年的时间里，晶体学一直是确定高分辨率生物分子结构的压倒性主导方法。来自单粒子低温电子显微镜和微电子衍射的竞争，对在低温下确定的结构中可能缺失或被破坏的功能相关信息的兴趣增加，以及对生物分子对化学和光刺激的响应的时间分辨研究的兴趣，推动了对室温下，更广泛地说，在从200 K附近的蛋白质-溶剂玻璃转变到∼350 K的温度下数据收集的新的兴趣。54，e1]。本文讨论了在非低温下采集结晶学数据的主要优点、物理原理和方法，以及与解释所得数据有关的一些因素。为了在结构生物学工具包中实现室温数据收集的潜力，应采用简化和标准化的方法，将家庭实验室准备的晶体送到同步加速器，并进行自动化处理和数据收集，类似于冰晶学的方法。
Key physical principles and methods for collecting crystallographic data from biomolecular systems at room temperature and, more generally, at temperatures between ∼200 and ∼350 K are reviewed and discussed. For roughly two decades, cryocrystallography has been the overwhelmingly dominant method for determining high-resolution biomolecular structures. Competition from single-particle cryo-electron microscopy and micro-electron diffraction, increased interest in functionally relevant information that may be missing or corrupted in structures determined at cryogenic temperature, and interest in time-resolved studies of the biomolecular response to chemical and optical stimuli have driven renewed interest in data collection at room temperature and, more generally, at temperatures from the protein–solvent glass transition near 200 K to ∼350 K. Fischer has recently reviewed practical methods for room-temperature data collection and analysis [Fischer (2021), Q. Rev. Biophys. 54, e1]. Here, the key advantages and physical principles of, and methods for, crystallographic data collection at noncryogenic temperatures and some factors relevant to interpreting the resulting data are discussed. For room-temperature data collection to realize its potential within the structural biology toolkit, streamlined and standardized methods for delivering crystals prepared in the home laboratory to the synchrotron and for automated handling and data collection, similar to those for cryocrystallography, should be implemented.
DOI：
10.1007/s10969-009-9074-y
发表时间：
2010-03
期刊：
Journal of structural and functional genomics
影响因子：
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作者：
Warkentin, Matthew;Thorne, Robert E
通讯作者：
Thorne, Robert E