Is Agarose an Impurity or an Impurity Filter? In Situ Observation of the Joint Gel/Impurity Effect on Protein Crystal Growth Kinetics
Is Agarose an Impurity or an Impurity Filter? In Situ Observation of the Joint Gel/Impurity Effect on Protein Crystal Growth Kinetics
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DOI：
10.1021/cg800157t
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发表时间：
2008-09
影响因子：
3.8
通讯作者：
A. V. Driessche;F. Otálora;J. Gavira;G. Sazaki
中科院分区：
文献类型：
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作者：
A. V. Driessche;F. Otálora;J. Gavira;G. Sazaki
通过比较从具有两种不同纯度水平（高度纯化（99.99%纯度）和商业级（98.5%纯度））的自由溶液和凝胶（琼脂糖）溶液中生长的晶体的二维（2D）成核速率和台阶速度，原位研究了琼脂糖凝胶和杂质对鸡蛋清溶菌酶晶体生长动力学的联合影响。使用激光共聚焦显微镜结合微分干涉对比显微镜（LCM - DIM）在四方溶菌酶晶体的{110}面上测量2D成核速率和台阶速度。作为异质成核位点的凝胶纤维的存在提高了2D成核速率。这些结果还表明，凝胶纤维/晶体界面和凝胶纤维/溶液界面的比表面能相似。这与观察到的琼脂糖纤维掺入溶菌酶晶格以及凝胶纤维对台阶速度的小影响是一致的。在凝胶和杂质同时存在的情况下的2D成核……
The joint effect of agarose gel and impurities on hen egg white lysozyme crystal growth kinetics was investigated in situ by comparing the two-dimensional (2D) nucleation rate and the step velocity of crystals growing from free and gelled (agarose) solutions having two different levels of purity: highly purified (99.99% pure) and commercial grade (98.5% pure). The 2D nucleation rate and step velocity were measured on {110} faces of tetragonal lysozyme crystals using laser confocal microscopy combined with differential interference contrast microscopy (LCM-DIM). 2D nucleation rates are enhanced by the presence of gel fibers that act as heterogeneous nucleation sites. These results also show that the specific surface energies are similar for the gel fiber/crystal interface and for the gel fiber/solution interface. This is consistent with the observed incorporation of agarose fibers into the lysozyme crystal lattice and the small effect of gel fibers on step velocity. 2D nucleation in the presence of both ge...