Selective Crystallization of Proteins Using Engineered Nanonucleants
Selective Crystallization of Proteins Using Engineered Nanonucleants
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DOI：
10.1021/cg201443s
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发表时间：
2012-03-01
影响因子：
3.8
通讯作者：
Heng, Jerry Y. Y.
中科院分区：
文献类型：
--
作者：
Shah, Umang V.;Williams, Daryl R.;Heng, Jerry Y. Y.
这项研究报告的第一次详细的实验研究的蛋白质结晶在工程nanofolined空间与控制孔径和窄孔径分布。我们提出了一个系统的方法来控制成核和结晶的生物大分子的蛋白质回转半径（R-g）和比孔径之间的关系的基础上。制备了一系列具有窄孔径分布的有序介孔的纳米核。测试模板的分子量范围为14至450 kDa的蛋白质。对于所有测试的蛋白质情况，仅在五种呈现的纳米成核剂中的一种上获得良好形成的蛋白质晶体，突出了这些成核剂所表现出的独特模板选择性。此外，伴刀豆球蛋白A和过氧化氢酶都结晶在类似的2倍低过饱和水平比以前报道的任何已知的方法。我们的观察结果完全支持理论研究，预测增强的热力学稳定性的蛋白质在nanocolinated腔，特别是成核剂孔径相对于蛋白质的回转半径的重要性。本文所述的成核剂可具有用于生物药物的下游分离和纯化的主要工业应用，以及用于结构测定的复杂蛋白质的结晶的改进的机会。
This study reports for the first time a detailed experimental investigation of protein crystallization in engineered nanoconfined spaces with both controlled pore diameters and narrow pore size distributions. We propose a systematic approach for controlling the nucleation and crystallization of biological macromolecules based on a relationship between the protein radius of gyration (R-g) and specific pore diameter. A series of nanonucleants with ordered mesopores having narrow pore size distributions were prepared. The templates were tested for proteins ranging in molecular weight from 14 to 450 kDa. Well-formed protein crystals were obtained on only one of the five presented nanonucleants for all protein cases tested, highlighting the unique template selectivity exhibited by these nucleants. In addition, concanavalin A and catalase were both crystallized at similar to 2 times lower supersaturation levels than previously reported by any known method. Our observations fully support theoretical studies that predict the enhanced thermodynamic stability of proteins in nanoconfined cavities, including specifically the importance of nucleant pore diameter with respect to protein radius of gyration. The nucleants described here could have major industrial applications for downstream separation and purification of biopharmaceuticals, as well as improved opportunities for the crystallization of complex proteins for structural determination.