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微通道中纳升尺度液滴内生长的蛋白质晶体的X射线衍射及其适用性评估。

X-ray diffraction of protein crystal grown in a nano-liter scale droplet in a microchannel and evaluation of its applicability.

作者信息

Maeki Masatoshi, Yoshizuka Saori, Yamaguchi Hiroshi, Kawamoto Masahide, Yamashita Kenichi, Nakamura Hiroyuki, Miyazaki Masaya, Maeda Hideaki

机构信息

Department of Molecular and Material Sciences, Interdisciplinary Graduate School of Engineering Sciences, Kyushu University, Kasuga-Koen, Kasuga, Fukuoka, Japan.

出版信息

Anal Sci. 2012;28(1):65. doi: 10.2116/analsci.28.65.

Abstract

We describe the technical aspects of the in-situ X-ray diffraction of a protein crystal prepared by a nanodroplet-based crystallization method. We were able to obtain diffraction patterns from a crystal grown in a capillary without any manipulation. Especially in our experimental approach, the crystals that moved to the nanodroplet interface were fixed strongly enough to carry out X-ray diffraction measurements that could be attributed to the high surface tension of the nanodroplet. The crystal was damaged by an indirect action of the X-rays because our in-situ X-ray diffraction measurement was carried out in the liquid phase without freezing the crystal; however, the obtained several diffraction patterns were of sufficiently fine quality for the crystal structure factors to be generated. We consider the technical examination presented in this paper to represent a seamless coupling of crystallization to X-ray analysis.

摘要

我们描述了通过基于纳米液滴的结晶方法制备的蛋白质晶体原位X射线衍射的技术细节。我们能够在不对晶体进行任何操作的情况下,从毛细管中生长的晶体获得衍射图谱。特别是在我们的实验方法中,移动到纳米液滴界面的晶体被牢固地固定住,足以进行X射线衍射测量,这可归因于纳米液滴的高表面张力。由于我们的原位X射线衍射测量是在液相中进行的,没有对晶体进行冷冻,晶体受到X射线的间接作用而受损;然而,所获得的几个衍射图谱的质量足够好,能够生成晶体结构因子。我们认为本文中提出的技术检验代表了结晶与X射线分析的无缝结合。

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