Fixed-target serial crystallography at the Structural Biology Center.

IF 2.4 3区物理与天体物理 Q2 INSTRUMENTS & INSTRUMENTATION

Journal of Synchrotron Radiation Pub Date : 2022-09-01 Epub Date: 2022-08-17 DOI:10.1107/S1600577522007895

Darren A Sherrell, Alex Lavens, Mateusz Wilamowski, Youngchang Kim, Ryan Chard, Krzysztof Lazarski, Gerold Rosenbaum, Rafael Vescovi, Jessica L Johnson, Chase Akins, Changsoo Chang, Karolina Michalska, Gyorgy Babnigg, Ian Foster, Andrzej Joachimiak

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Abstract

Serial synchrotron crystallography enables the study of protein structures under physiological temperature and reduced radiation damage by collection of data from thousands of crystals. The Structural Biology Center at Sector 19 of the Advanced Photon Source has implemented a fixed-target approach with a new 3D-printed mesh-holder optimized for sample handling. The holder immobilizes a crystal suspension or droplet emulsion on a nylon mesh, trapping and sealing a near-monolayer of crystals in its mother liquor between two thin Mylar films. Data can be rapidly collected in scan mode and analyzed in near real-time using piezoelectric linear stages assembled in an XYZ arrangement, controlled with a graphical user interface and analyzed using a high-performance computing pipeline. Here, the system was applied to two β-lactamases: a class D serine β-lactamase from Chitinophaga pinensis DSM 2588 and L1 metallo-β-lactamase from Stenotrophomonas maltophilia K279a.

Abstract Image

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结构生物学中心的固定目标序列晶体学。

串行同步加速器晶体学能够在生理温度下研究蛋白质结构，并通过收集成千上万晶体的数据来减少辐射损伤。位于先进光子源 19 区的结构生物学中心采用了一种固定目标的方法，这种方法使用了一种新的三维打印网状支架，优化了样品处理。该支架将晶体悬浮液或液滴乳液固定在尼龙网上，在两层薄薄的 Mylar 膜之间的母液中捕获并密封近单层晶体。数据可以在扫描模式下快速收集，并通过以 XYZ 排列方式组装的压电线性平台进行近乎实时的分析，这些数据由图形用户界面控制，并通过高性能计算管道进行分析。在这里，该系统被应用于两种 β-内酰胺酶：来自 Chitinophaga pinensis DSM 2588 的 D 类丝氨酸 β-内酰胺酶和来自 Stenotrophomonas maltophilia K279a 的 L1 金属-β-内酰胺酶。

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来源期刊

Journal of Synchrotron Radiation 物理-光学

CiteScore

5.10

自引率

12.00%

发文量

289

审稿时长

4-8 weeks

期刊介绍： Synchrotron radiation research is rapidly expanding with many new sources of radiation being created globally. Synchrotron radiation plays a leading role in pure science and in emerging technologies. The Journal of Synchrotron Radiation provides comprehensive coverage of the entire field of synchrotron radiation and free-electron laser research including instrumentation, theory, computing and scientific applications in areas such as biology, nanoscience and materials science. Rapid publication ensures an up-to-date information resource for scientists and engineers in the field.