A linear relationship between crystal size and fragment binding time observed crystallographically: implications for fragment library screening using acoustic droplet ejection.
A linear relationship between crystal size and fragment binding time observed crystallographically: implications for fragment library screening using acoustic droplet ejection.
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DOI：
10.1371/journal.pone.0101036
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Soares AS
中科院分区：
文献类型：
--
作者：
Cole K;Roessler CG;Mulé EA;Benson-Xu EJ;Mullen JD;Le BA;Tieman AM;Birone C;Brown M;Hernandez J;Neff S;Williams D;Allaire M;Orville AM;Sweet RM;Soares AS
高通量筛选技术，如声学液滴喷射（ADE），大大提高了从晶体中获取X射线衍射数据的速度。ADE的一个有前景的高通量筛选应用是快速将蛋白质晶体与片段文库联合收割机组合。在这种方法中，每个片段直接在数据收集介质上或在移动的传送带上浸入蛋白质晶体，然后将晶体传送到X射线束。通过同时处理多个晶体与碎片标本相结合，这些技术放松了自动计数器占空比瓶颈，目前阻碍了第三代同步加速器的最佳利用。两个因素限制了适合使用ADE等技术进行片段筛选的项目的速度和范围。首先，在高通量筛选设备位于X射线站（例如上述传送带系统）内部的应用中，数据采集的速度受到每个碎片浸入其蛋白质晶体所需的时间的限制。其次，在晶体与片段直接在数据采集介质上组合的应用中（包括上述两种ADE方法），片段必须浸泡到晶体中的最大时间受到片段浸泡期间蛋白质晶体的蒸发脱水的限制。在这里，我们证明，这两个问题可以通过使用小晶体最小化，因为浸泡时间所需的碎片命中，以达到高占有率依赖于晶体尺寸近似线性。
High throughput screening technologies such as acoustic droplet ejection (ADE) greatly increase the rate at which X-ray diffraction data can be acquired from crystals. One promising high throughput screening application of ADE is to rapidly combine protein crystals with fragment libraries. In this approach, each fragment soaks into a protein crystal either directly on data collection media or on a moving conveyor belt which then delivers the crystals to the X-ray beam. By simultaneously handling multiple crystals combined with fragment specimens, these techniques relax the automounter duty-cycle bottleneck that currently prevents optimal exploitation of third generation synchrotrons. Two factors limit the speed and scope of projects that are suitable for fragment screening using techniques such as ADE. Firstly, in applications where the high throughput screening apparatus is located inside the X-ray station (such as the conveyor belt system described above), the speed of data acquisition is limited by the time required for each fragment to soak into its protein crystal. Secondly, in applications where crystals are combined with fragments directly on data acquisition media (including both of the ADE methods described above), the maximum time that fragments have to soak into crystals is limited by evaporative dehydration of the protein crystals during the fragment soak. Here we demonstrate that both of these problems can be minimized by using small crystals, because the soak time required for a fragment hit to attain high occupancy depends approximately linearly on crystal size.
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DOI：
10.1107/s0907444906012601
发表时间：
2006-06-01
期刊：
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY
影响因子：
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发表时间：
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影响因子：
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发表时间：
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影响因子：
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