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高速固定靶串行病毒晶体学

High-speed fixed-target serial virus crystallography.

作者信息

Roedig Philip, Ginn Helen M, Pakendorf Tim, Sutton Geoff, Harlos Karl, Walter Thomas S, Meyer Jan, Fischer Pontus, Duman Ramona, Vartiainen Ismo, Reime Bernd, Warmer Martin, Brewster Aaron S, Young Iris D, Michels-Clark Tara, Sauter Nicholas K, Kotecha Abhay, Kelly James, Rowlands David J, Sikorsky Marcin, Nelson Silke, Damiani Daniel S, Alonso-Mori Roberto, Ren Jingshan, Fry Elizabeth E, David Christian, Stuart David I, Wagner Armin, Meents Alke

机构信息

Photon Science, Deutsches Elektronen-Synchrotron DESY, Hamburg, Germany.

Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK.

出版信息

Nat Methods. 2017 Aug;14(8):805-810. doi: 10.1038/nmeth.4335. Epub 2017 Jun 19.

Abstract

We report a method for serial X-ray crystallography at X-ray free-electron lasers (XFELs), which allows for full use of the current 120-Hz repetition rate of the Linear Coherent Light Source (LCLS). Using a micropatterned silicon chip in combination with the high-speed Roadrunner goniometer for sample delivery, we were able to determine the crystal structures of the picornavirus bovine enterovirus 2 (BEV2) and the cytoplasmic polyhedrosis virus type 18 polyhedrin, with total data collection times of less than 14 and 10 min, respectively. Our method requires only micrograms of sample and should therefore broaden the applicability of serial femtosecond crystallography to challenging projects for which only limited sample amounts are available. By synchronizing the sample exchange to the XFEL repetition rate, our method allows for most efficient use of the limited beam time available at XFELs and should enable a substantial increase in sample throughput at these facilities.

摘要

我们报告了一种在X射线自由电子激光(XFEL)上进行串行X射线晶体学分析的方法,该方法能够充分利用线性相干光源(LCLS)当前120赫兹的重复频率。通过将微图案化硅芯片与用于样品输送的高速“走鹃”测角仪相结合,我们能够分别在总数据收集时间少于14分钟和10分钟的情况下,确定微小核糖核酸病毒牛肠道病毒2(BEV2)和18型细胞质多角体病毒多角体蛋白的晶体结构。我们的方法仅需要微克级别的样品,因此应能将串行飞秒晶体学的适用性扩展到那些仅有少量样品可用的具有挑战性的项目中。通过将样品交换与XFEL重复频率同步,我们的方法能够最有效地利用XFEL有限的束流时间,并应能大幅提高这些设施的样品通量。

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High-speed fixed-target serial virus crystallography.高速固定靶串行病毒晶体学
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本文引用的文献

1
Low-dose fixed-target serial synchrotron crystallography.低剂量固定靶标串行同步辐射晶体学。
Acta Crystallogr D Struct Biol. 2017 Apr 1;73(Pt 4):373-378. doi: 10.1107/S2059798317002996. Epub 2017 Mar 31.
4
On the release of for processing X-ray free-electron laser images.关于用于处理X射线自由电子激光图像的[具体内容缺失]的发布。
J Appl Crystallogr. 2016 May 11;49(Pt 3):1065-1072. doi: 10.1107/S1600576716006981. eCollection 2016 Jun 1.
7
High-density grids for efficient data collection from multiple crystals.用于从多个晶体高效收集数据的高密度网格。
Acta Crystallogr D Struct Biol. 2016 Jan;72(Pt 1):2-11. doi: 10.1107/S2059798315020847. Epub 2016 Jan 1.
10
Serial femtosecond X-ray diffraction of enveloped virus microcrystals.包埋病毒微晶的连续飞秒 X 射线衍射。
Struct Dyn. 2015 Aug 20;2(4):041720. doi: 10.1063/1.4929410. eCollection 2015 Jul.

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