A simple and rapid method for mounting protein crystals at room temperature
A simple and rapid method for mounting protein crystals at room temperature
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一种简单快速的室温安装蛋白质晶体的方法
DOI：
10.1107/s0021889802019623
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
A. D'arcy
中科院分区：
文献类型：
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作者：
A. M. Sweeney;A. D'arcy
用于X射线数据采集的蛋白质晶体冷冻现已成为大多数生物结构实验室的常规方法。同步加速器设施的改进及其使用量的增加使得适当地冷冻晶体对于最佳数据收集是必不可少的。虽然在一般情况下，晶体可以在不造成重大损害的情况下冷却，但在一些情况下，具有轻微无序或孪生问题的晶体在冻结过程中会遭受相当大的损失。在其他情况下，不良或马赛克衍射可能归咎于冷冻保护剂或冷却方案。当晶体的固有质量较差时，许多晶体可能会浪费在寻找最佳冷冻条件的过程中。原则上，收集室温衍射数据将提供一个参考，以便能够检测添加低温保护剂或低温冷却造成的晶体损伤。然而，在实践中，许多研究人员不愿这样做，原因之一是毛细管安装晶体是一种繁琐的方法，特别是对那些对结晶学新手来说。本文报道了一种在室温下安装晶体的简化方法，该方法不需要太多的专业知识，也不需要昂贵的设备。
Cryocooling of protein crystals for X-ray data collection has now become a routine method in the majority of biostructural laboratories. The improvement of facilities at synchrotron sources and their increased use has made it essential to have properly frozen crystals for optimal data collection. Although in general crystals can be cooled without significant damage, there are often cases in which crystals with slight disorder or twinning problems suffer considerably during the freezing process. In other cases, poor or mosaic diffraction may be blamed on the cryoprotectant or cooling protocol. Many crystals may be wasted in searching for the best freezing conditions when the intrinsic quality of the crystals is poor. In principle, the collection of room-temperature diffraction data would provide a reference that would allow the detection of crystal damage caused by addition of cryoprotectant or by cryocooling. In practice, however, many investigators are reluctant to do this, one reason being that capillary mounting of crystals is a tedious method, especially for those who are new to crystallography. Here a simplified method for mounting crystals at room temperature is reported, which requires little expertise and no expensive equipment.