采用连续同步辐射晶体学进行时间分辨分析的液体应用方法。

Liquid application method for time-resolved analyses by serial synchrotron crystallography.

机构信息

Max-Planck-Institute for Structure and Dynamics of Matter, Department for Atomically Resolved Dynamics, Hamburg, Germany.

Centre for Ultrafast Imaging, Universität Hamburg, Hamburg, Germany.

出版信息

Nat Methods. 2019 Oct;16(10):979-982. doi: 10.1038/s41592-019-0553-1. Epub 2019 Sep 16.

DOI:10.1038/s41592-019-0553-1

PMID:31527838

Abstract

We introduce a liquid application method for time-resolved analyses (LAMA), an in situ mixing approach for serial crystallography. Picoliter-sized droplets are shot onto chip-mounted protein crystals, achieving near-full ligand occupancy within theoretical diffusion times. We demonstrate proof-of-principle binding of GlcNac to lysozyme, and resolve glucose binding and subsequent ring opening in a time-resolved study of xylose isomerase.

摘要

我们介绍了一种用于时间分辨分析的液相应用方法（LAMA），这是一种用于连续结晶学的原位混合方法。皮升级大小的液滴滴到芯片上的蛋白质晶体上，在理论扩散时间内实现了接近完全的配体占据。我们通过 GlcNac 与溶菌酶的结合证明了这一原理，并且在木糖异构酶的时间分辨研究中观察到了葡萄糖结合和随后的环打开。