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Surfaces with Controllable Topography and Chemistry Used as a Template for Protein Crystallization

机译：具有可控地形和化学的表面用作蛋白质结晶的模板

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摘要

Surfaces with controllable topography and chemistry were prepared to act as substrates for protein crystallization, in order to investigate the influence of these surface properties on the protein crystallization outcome. Three different methods were investigated to deposit 1,3,5-tris(10-carboxydecyloxy)benzene (TCDB) on a muscovite mica substrate to find the best route for controlled topography. Of these three, sublimation worked best. Contact angle measurements revealed that the surfaces with short exposure to the TCDB vapor (20 min or less) are hydrophilic, while surfaces exposed for 30 min or longer are hydrophobic. The hydrophilic surfaces are flat with low steps, while the hydrophobic surfaces contain macrosteps. Four model proteins were used for crystallization on the surfaces with controlled topography and chemistry. Hen egg white lysozyme crystals were less numerous on the surface with macrosteps than on smoother surfaces. On the other hand, insulin nucleated faster on the hydrophobic surfaces with macrosteps, and therefore, the crystals were more abundant and smaller. Bovine serum albumin and talin protein crystals were more numerous on all TCDB functionalized surfaces, compared to the reference clean muscovite mica surfaces. Overall, this shows that surface topography and chemistry is an important factor that partly determines the outcome in a protein crystallization experiment.
机译：制备具有可控地形和化学性的表面以用作蛋白质结晶的基材，以研究这些表面性质对蛋白质结晶结果的影响。研究了三种不同的方法，将1,3,5-三（10-羧基氧基）苯（TCDB）沉积在Muscovite MICA底物上，以找到受控地形的最佳途径。在这三个中，升华效果最佳。接触角测量表明，具有短路暴露于TCDB蒸气（20分钟或更短）的表面是亲水的，而暴露30分钟或更长的表面是疏水的。亲水性表面与低步骤平坦，而疏水性表面含有大麦纤维菌。使用四种模型蛋白质在具有受控形貌和化学的表面上结晶。母鸡蛋白溶菌酶晶体较少，在表面上具有大鼠比在更平滑的表面上的表面上。另一方面，胰岛素在疏水表面上含有大麦粥样物的疏水表面，因此，晶体更加丰富和更小。与参考清洁的Muscovite云母表面相比，所有TCDB官能化表面上牛血清白蛋白和少素蛋白质晶体更加多。总体而言，这表明表面形貌和化学是部分决定蛋白质结晶实验中的结果的重要因素。

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  • 来源
    《Crystal growth & design》 |2018年第2期|共7页
  • 作者

    Wester de Poel; Joris A. W. Münninghoff; Johannes A. A. W. Elemans; Willem J. P. van Enckevort; Alan E. Rowan; Elias Vlieg;

  • 作者单位

    Radboud University Institute for Molecules and Materials Heyendaalseweg 135 Nijmegen 6525 AJ The Netherlands;

    Radboud University Institute for Molecules and Materials Heyendaalseweg 135 Nijmegen 6525 AJ The Netherlands;

    Radboud University Institute for Molecules and Materials Heyendaalseweg 135 Nijmegen 6525 AJ The Netherlands;

    Radboud University Institute for Molecules and Materials Heyendaalseweg 135 Nijmegen 6525 AJ The Netherlands;

    Radboud University Institute for Molecules and Materials Heyendaalseweg 135 Nijmegen 6525 AJ The Netherlands;

    Radboud University Institute for Molecules and Materials Heyendaalseweg 135 Nijmegen 6525 AJ The Netherlands;

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  • 正文语种 eng
  • 中图分类 晶体学;
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