Rapid and efficient room-temperature serial synchrotron crystallography using the CFEL TapeDrive.
Rapid and efficient room-temperature serial synchrotron crystallography using the CFEL TapeDrive.
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使用CFEL磁带驱动器，最大限度地减少样品消耗和数据收集时间所需的串行同步辐射晶体学（SSX）实现和第一个室温结构的两种酶的描述，以及评估的好处SSX的辐射敏感蛋白质。在传统的同步加速器光源（SSX）的连续晶体学提供了在室温下使用生物大分子的微米级晶体常规收集数据的可能性。然而，SSX数据收集还不是常规的，目前需要比标准的旋转系列低温结晶学更长的时间。因此，其用于高通量方法，如基于片段的药物筛选，其中在生理学温度下测量的可能性将是一个很大的好处，是受损的。在使用基于传送带的样品递送系统-CFEL TapeDrive -与三种不同的生物相关蛋白质（肺炎克雷伯氏菌CTX-M-14 β-内酰胺酶、红球菌木聚糖酶GH 11和黄曲霉尿酸氧化酶）进行高通量SSX的过程中，这里显示可以在不到一分钟的时间内收集完整的数据集，并且只需要最少量的样品。
Using the CFEL TapeDrive, minimization of sample consumption and data collection times needed for serial synchrotron crystallography (SSX) were achieved and the first room-temperature structures of two enzymes are described as well as an assessment of the benefit of SSX for a radiation-sensitive protein. Serial crystallography at conventional synchrotron light sources (SSX) offers the possibility to routinely collect data at room temperature using micrometre-sized crystals of biological macromolecules. However, SSX data collection is not yet as routine and currently takes significantly longer than the standard rotation series cryo-crystallography. Thus, its use for high-throughput approaches, such as fragment-based drug screening, where the possibility to measure at physiological temperatures would be a great benefit, is impaired. On the way to high-throughput SSX using a conveyor belt based sample delivery system – the CFEL TapeDrive – with three different proteins of biological relevance (Klebsiella pneumoniae CTX-M-14 β-lactamase, Nectria haematococca xylanase GH11 and Aspergillus flavus urate oxidase), it is shown here that complete datasets can be collected in less than a minute and only minimal amounts of sample are required.