Fabrication of X-ray compatible microfluidic platforms for protein crystallization.
Fabrication of X-ray compatible microfluidic platforms for protein crystallization.
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DOI：
10.1016/j.snb.2012.08.048
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发表时间：
2012-11
影响因子：
8.4
通讯作者：
Kenis, Paul J. A.
中科院分区：
文献类型：
--
作者：
Guha, Sudipto;Perry, Sarah L.;Pawate, Ashtamurthy S.;Kenis, Paul J. A.
关键词：
本文报道了一种利用不同材料制作多层微流控蛋白质结晶平台的方法，以实现X射线的透明性和与结晶剂的相容性。为了验证这种方法，我们在芯片上结晶了溶菌酶、苏维菌素和核糖核酸酶A这三种可溶性蛋白质，然后进行了芯片上的衍射数据采集。我们还报告了一种带有孔阵列的芯片，与传统的筛选方法相比，该芯片用于筛选消耗最少蛋白质溶液的不同条件。在井中可以生长出大量高质量的同晶蛋白质晶体，之后可以从仍然驻留在井中的许多晶体中收集X射线切片数据。通过合并这些切片的数据，然后用结晶学软件进行进一步处理，可以获得完整的蛋白质结构。这种使用X射线透明芯片进行筛选、晶体生长和X射线数据收集的方法能够从并行安装的多个晶体中收集室温数据，从而消除了对晶体的处理并将对晶体的辐射损害降至最低。
This paper reports a method for fabricating multilayer microfluidic protein crystallization platforms using different materials to achieve X-ray transparency and compatibility with crystallization reagents. To validate this approach, three soluble proteins, lysozyme, thaumatin, and ribonuclease A were crystallized on-chip, followed by on-chip diffraction data collection. We also report a chip with an array of wells for screening different conditions that consume a minimal amount of protein solution as compared to traditional screening methods. A large number of high quality isomorphous protein crystals can be grown in the wells, after which slices of X-ray data can be collected from many crystals still residing within the wells. Complete protein structures can be obtained by merging these slices of data followed by further processing with crystallography software. This approach of using an x-ray transparent chip for screening, crystal growth, and X-ray data collection enables room temperature data collection from many crystals mounted in parallel, which thus eliminates crystal handling and minimizes radiation damage to the crystals.
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影响因子：
3.4
作者：
Liu, Wei;Hanson, Michael A.;Cherezov, Vadim
通讯作者：
Cherezov, Vadim
DOI：
10.1107/s0907444911011589
发表时间：
2011-06
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Kisselman G;Qiu W;Romanov V;Thompson CM;Lam R;Battaile KP;Pai EF;Chirgadze NY
通讯作者：
Chirgadze NY
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
64.8
作者：
CHUNG, H;CAFFREY, M
通讯作者：
CAFFREY, M
影响因子：
3.8
作者：
Caffrey, Martin
通讯作者：
Caffrey, Martin