The plug-based nanovolume Microcapillary Protein Crystallization System (MPCS).
The plug-based nanovolume Microcapillary Protein Crystallization System (MPCS).
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DOI：
10.1107/s0907444908028060
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发表时间：
2008-11
期刊：
影响因子：
--
通讯作者：
Stewart L
中科院分区：
文献类型：
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作者：
Gerdts CJ;Elliott M;Lovell S;Mixon MB;Napuli AJ;Staker BL;Nollert P;Stewart L
微毛细管蛋白质结晶系统（MPCS）是一种新的蛋白质结晶技术，用于产生纳米级的结晶实验，用于晶体筛选和优化。使用MPCS，在塑料MPCS CrystalCard中生长衍射就绪的晶体，并用于解析甲硫氨酸-R-亚砜还原酶的结构。微毛细管蛋白质结晶系统（MPCS）体现了一种新的半自动化基于插件的结晶技术，该技术能够以塑料器皿的形式对结晶条件进行纳升体积筛选，从而可以轻松地去除晶体以进行传统的冷冻保护和X射线衍射数据收集。在这些塑料装置中生长的蛋白质晶体可以直接进行原位X射线衍射研究。MPCS将结晶混合物的配制与结晶实验的准备相结合。在塑料CrystalCard的微流控Teflon管或微流控回路内，产生约10-20 nl体积的液滴，每个液滴代表具有不同化学组成的微批式结晶实验。 整个蛋白质样品用于结晶实验。稀疏矩阵筛选和化学梯度筛选可以结合在一个全面的“混合”结晶试验。该技术适合于通过使用优化试剂如沉淀剂、配体或冷冻保护剂的高粒度梯度筛选进行优化。
The Microcapillary Protein Crystallization System (MPCS) is a new protein-crystallization technology used to generate nanolitre-sized crystallization experiments for crystal screening and optimization. Using the MPCS, diffraction-ready crystals were grown in the plastic MPCS CrystalCard and were used to solve the structure of methionine-R-sulfoxide reductase. The Microcapillary Protein Crystallization System (MPCS) embodies a new semi-automated plug-based crystallization technology which enables nanolitre-volume screening of crystallization conditions in a plasticware format that allows crystals to be easily removed for traditional cryoprotection and X-ray diffraction data collection. Protein crystals grown in these plastic devices can be directly subjected to in situ X-ray diffraction studies. The MPCS integrates the formulation of crystallization cocktails with the preparation of the crystallization experiments. Within microfluidic Teflon tubing or the microfluidic circuitry of a plastic CrystalCard, ∼10–20 nl volume droplets are generated, each representing a microbatch-style crystallization experiment with a different chemical composition. The entire protein sample is utilized in crystallization experiments. Sparse-matrix screening and chemical gradient screening can be combined in one comprehensive ‘hybrid’ crystallization trial. The technology lends itself well to optimization by high-granularity gradient screening using optimization reagents such as precipitation agents, ligands or cryoprotectants.