Weik Martin, Colletier Jacques Philippe
CEA, IBS, Laboratoire de Biophysique Moléculaire, F-38054 Grenoble, France.
Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):437-46. doi: 10.1107/S0907444910002702. Epub 2010 Mar 24.
X-ray crystallography provides structural details of biological macromolecules. Whereas routine data are collected close to 100 K in order to mitigate radiation damage, more exotic temperature-controlled experiments in a broader temperature range from 15 K to room temperature can provide both dynamical and structural insights. Here, the dynamical behaviour of crystalline macromolecules and their surrounding solvent as a function of cryo-temperature is reviewed. Experimental strategies of kinetic crystallography are discussed that have allowed the generation and trapping of macromolecular intermediate states by combining reaction initiation in the crystalline state with appropriate temperature profiles. A particular focus is on recruiting X-ray-induced changes for reaction initiation, thus unveiling useful aspects of radiation damage, which otherwise has to be minimized in macromolecular crystallography.
X射线晶体学可提供生物大分子的结构细节。常规数据采集通常在接近100 K的温度下进行,以减轻辐射损伤,而在从15 K到室温的更宽温度范围内进行的更特殊的温控实验,能够提供动力学和结构方面的见解。本文综述了晶体大分子及其周围溶剂在低温下的动力学行为。讨论了动力学晶体学的实验策略,这些策略通过将晶体状态下的反应引发与适当的温度分布相结合,实现了大分子中间态的生成与捕获。特别关注利用X射线诱导的变化来引发反应,从而揭示辐射损伤的有用方面,而在大分子晶体学中,辐射损伤通常需要被降至最低。