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Tailored Hydrogel Membranes for Efficient Protein Crystallization

机译：量身定制的水凝胶膜，可实现高效的蛋白质结晶

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摘要

Crystallization still represents the bottleneck in the process of protein structure determination at high resolution, despite high-throughput structural genomics programs requiring optimized crystallization strategies regarding crystal quality, time, success rate, reproducibility, and used protein amount. On the other hand, the development of suitable materials for controlled heterogeneous nucleation might facilitate biomacromolecular crystallization in a variety of experimental conditions which are not conventionally fruitful. Here, the possibility to fabricate hydrogel membranes displaying controlled chemical composition and nanostructure and to use them as heterogeneous supports for biomacromolecular crystallization is demonstrated. Diverse gel morphologies are obtained by controlling phase separation kinetics during gel layer formation on membrane support. These composite materials are found to increase the efficiency of the crystallization process so that crystals with enhanced diffraction properties are produced at lower protein concentration than conventional technique, thus affording the possibility to improve current approaches to protein crystallization and to be adapted to specific targets.
机译：尽管高通量结构基因组学程序需要有关晶体质量，时间，成功率，可再现性和已用蛋白质量的优化结晶策略，但结晶仍代表着高分辨率蛋白质结构确定过程中的瓶颈。另一方面，开发用于受控异质成核的合适材料可能会促进生物大分子结晶，而这些条件在常规上并不富有成效的各种实验条件下都可实现。在这里，证明了制造具有可控化学成分和纳米结构的水凝胶膜并将其用作生物大分子结晶的异质载体的可能性。通过控制在膜载体上形成凝胶层期间的相分离动力学，可以获得不同的凝胶形态。发现这些复合材料提高了结晶过程的效率，从而在比常规技术更低的蛋白质浓度下产生具有增强的衍射特性的晶体，从而提供了改善当前蛋白质结晶方法并适应特定目标的可能性。

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  • 来源
    《Advanced Functional Materials》 |2014年第11期|1582-1590|共9页
  • 作者

    Gianluca Di Profio; Marietta Polino; Fiore P. Nicoletta; Benny D. Behiso; Rocco Caliandro; Enrica Fontananova; Giovanni De Filpo; Efrem Curcio; Enrico Drioli;

  • 作者单位

    National Research Council of Italy (CNR) Institute on Membrane Technology (ITM) Via P. Bucci c/o Universita della Calabria Cubo 17/C, 87036 Rende (CS), Italy;

    University of Calabria Department of Pharmacy Health and Nutritional Sciences Via P. Bucci, Ed., Polifunzionale, 87036 Rende (CS), Italy;

    University of Calabria Department of Chemistry and Chemical Technologies Via P. Bucci Cubo 15/C, 87036 Rende (CS), Italy;

    National Research Council of Italy (CNR) Institute of Crystallography (IC) Via G. Amendola 122/o, 70125 Bari (BA), Italy;

    National Research Council of Italy (CNR) Institute of Crystallography (IC) Via G. Amendola 122/o, 70125 Bari (BA), Italy;

    National Research Council of Italy (CNR) Institute on Membrane Technology (ITM) Via P. Bucci c/o Universita della Calabria Cubo 17/C, 87036 Rende (CS), Italy;

    University of Calabria Department of Chemistry and Chemical Technologies Via P. Bucci Cubo 15/C, 87036 Rende (CS), Italy;

    University of Calabria Department of Environmental and Chemical Engineering Via P. Bucci Cubo 15/C, 87036 Rende (CS), Italy;

    University of Calabria Department of Environmental and Chemical Engineering Via P. Bucci Cubo 15/C, 87036 Rende (CS), Italy;

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