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高通量微晶递送系统在串行飞秒晶体学中的应用。

Application of a high-throughput microcrystal delivery system to serial femtosecond crystallography.

作者信息

Lee Donghyeon, Park Sehan, Lee Keondo, Kim Jangwoo, Park Gisu, Nam Ki Hyun, Baek Sangwon, Chung Wan Kyun, Lee Jong-Lam, Cho Yunje, Park Jaehyun

机构信息

Department of Mechanical Engineering, POSTECH, 77 Cheongam-Ro, Pohang, 37673, Republic of Korea.

PAL-XFEL, Pohang Accelerator Laboratory, 80 Jigok-ro 127 beongil, Pohang, 37673, Republic of Korea.

出版信息

J Appl Crystallogr. 2020 Mar 25;53(Pt 2):477-485. doi: 10.1107/S1600576720002423. eCollection 2020 Apr 1.

DOI:10.1107/S1600576720002423
PMID:32280322
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7133064/
Abstract

Microcrystal delivery methods are pivotal in the use of serial femtosecond crystallography (SFX) to resolve the macromolecular structures of proteins. Here, the development of a novel technique and instruments for efficiently delivering microcrystals for SFX are presented. The new method, which relies on a one-dimensional fixed-target system that includes a microcrystal container, consumes an extremely low amount of sample compared with conventional two-dimensional fixed-target techniques at ambient temperature. This novel system can deliver soluble microcrystals without highly viscous carrier media and, moreover, can be used as a microcrystal growth device for SFX. Diffraction data collection utilizing this advanced technique along with a real-time visual servo scan system has been successfully demonstrated for the structure determination of proteinase K microcrystals at 1.85 Å resolution.

摘要

微晶递送方法在使用串行飞秒晶体学（SFX）解析蛋白质的大分子结构中起着关键作用。本文介绍了一种用于高效递送微晶以进行SFX的新技术和仪器的开发。这种新方法依赖于一个包含微晶容器的一维固定靶系统，与传统的二维固定靶技术相比，在环境温度下消耗的样品量极低。这种新型系统可以在没有高粘性载体介质的情况下递送可溶性微晶，此外，还可以用作SFX的微晶生长装置。利用这种先进技术结合实时视觉伺服扫描系统进行的衍射数据收集，已成功用于分辨率为1.85 Å的蛋白酶K微晶的结构测定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7a1/7133064/f064604bd939/j-53-00477-fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7a1/7133064/d9889586498c/j-53-00477-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7a1/7133064/4d828329ff1f/j-53-00477-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7a1/7133064/e34e8db22152/j-53-00477-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7a1/7133064/4cebb758d406/j-53-00477-fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7a1/7133064/f064604bd939/j-53-00477-fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7a1/7133064/d9889586498c/j-53-00477-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7a1/7133064/4d828329ff1f/j-53-00477-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7a1/7133064/e34e8db22152/j-53-00477-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7a1/7133064/4cebb758d406/j-53-00477-fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7a1/7133064/f064604bd939/j-53-00477-fig5.jpg

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