Humidity control and hydrophilic glue coating applied to mounted protein crystals improves X-ray diffraction experiments
Humidity control and hydrophilic glue coating applied to mounted protein crystals improves X-ray diffraction experiments
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DOI：
10.1107/s0907444913018027
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发表时间：
2013-09-01
影响因子：
2.2
通讯作者：
Kumasaka, Takashi
中科院分区：
文献类型：
--
作者：
Baba, Seiki;Hoshino, Takeshi;Kumasaka, Takashi
蛋白质晶体是易碎的，有时很难在进行X射线衍射实验之前找到适合处理和低温冷却晶体的条件。为了克服这个问题，已经开发了一种蛋白质晶体安装方法，该方法涉及水溶性聚合物和可以调节安装晶体的水分含量的受控潮湿空气。通过用聚合物胶涂覆晶体并将其暴露于受控的潮湿空气中，晶体在室温下稳定，并在优化的湿度下低温冷却。此外，胶涂覆的晶体可重复地显示出其晶格常数随湿度变化的逐渐转变;因此，使用这种方法，可以制备一系列同晶晶体。这项技术在研究脆弱的蛋白质晶体（包括膜蛋白）时很有价值，也将有助于多晶体数据收集。
Protein crystals are fragile, and it is sometimes difficult to find conditions suitable for handling and cryocooling the crystals before conducting X-ray diffraction experiments. To overcome this issue, a protein crystal-mounting method has been developed that involves a water-soluble polymer and controlled humid air that can adjust the moisture content of a mounted crystal. By coating crystals with polymer glue and exposing them to controlled humid air, the crystals were stable at room temperature and were cryocooled under optimized humidity. Moreover, the glue-coated crystals reproducibly showed gradual transformations of their lattice constants in response to a change in humidity; thus, using this method, a series of isomorphous crystals can be prepared. This technique is valuable when working on fragile protein crystals, including membrane proteins, and will also be useful for multi-crystal data collection.