Millisecond mix-and-quench crystallography (MMQX) enables time-resolved studies of PEPCK with remote data collection.
Millisecond mix-and-quench crystallography (MMQX) enables time-resolved studies of PEPCK with remote data collection.
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DOI：
10.1107/s2052252521007053
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发表时间：
2021-09-01
期刊：
影响因子：
3.9
通讯作者：
Thorne RE
中科院分区：
文献类型：
--
作者：
Clinger JA;Moreau DW;McLeod MJ;Holyoak T;Thorne RE
低温捕获技术的改进使酶的单晶时间分辨晶体学具有40 ms的时间分辨率。 随着连续晶体学方法的改进，生物分子的时间分辨晶体学在行动中迅速发展，但所需的大量晶体和复杂的实验基础设施仍然是其广泛应用的严重障碍。在这里，毫秒混合和淬火晶体学（MMQX）已经开发出来，它产生毫秒时间分辨的数据，使用少得多的晶体和常规远程同步数据收集。为了证明MMQX的能力，以40 ms的时间分辨率观察到磷酸烯醇丙酮酸羧基激酶（PEPCK）将草酰乙酸转化为磷酸烯醇丙酮酸。 通过降低时间分辨晶体学的进入门槛，MMQX应该能够广泛扩展蛋白质动力学的结构研究。
Improvements in cryo-trapping technology enable single-crystal time-resolved crystallography of an enzyme with 40 ms time resolution. Time-resolved crystallography of biomolecules in action has advanced rapidly as methods for serial crystallography have improved, but the large number of crystals and the complex experimental infrastructure that are required remain serious obstacles to its widespread application. Here, millisecond mix-and-quench crystallography (MMQX) has been developed, which yields millisecond time-resolved data using far fewer crystals and routine remote synchrotron data collection. To demonstrate the capabilities of MMQX, the conversion of oxaloacetic acid to phosphoenolpyruvate by phosphoenolpyruvate carboxykinase (PEPCK) is observed with a time resolution of 40 ms. By lowering the entry barrier to time-resolved crystallography, MMQX should enable a broad expansion in structural studies of protein dynamics.