Nucleation and Growth Kinetics in Protein Crystallization

一年来取得的成绩如下：日本组（j组）采用双光束干涉法测定了蛋清溶菌酶（HEW溶菌酶）的四方型、正交型和三斜型溶菌酶的溶解度。J-group开发了一种干涉测量晶体（HEW溶菌酶）内部折射率的方法。j组采用高效液相色谱法获得高纯度HEW溶菌酶样品。有了这个样本，他们用原子力显微镜观察了晶体界面。与美国组（a组）的结果进行对比讨论。j组尝试在高磁场条件下对HEW溶菌酶进行结晶。他们发现它可以控制晶体的成核速率和取向。apap铁蛋白（a组）和铁蛋白（j组）经高纯度纯化后结晶。他们再次证明了杂质的去除对于高质量晶体的生长是必不可少的。j组形成铁蛋白形态。a组对HEW溶菌酶溶液进行了光散射和多环实验，并讨论了其聚集过程。他们估计了溶液的二次维里系数，并讨论了离子强度与分子间相互作用的关系。a组利用激光干涉测量法对邻面形貌进行精确测量，得到邻面法向、阶跃生长速率和斜率随时间的变化。a组发现HEW溶菌酶溶液存在一个区域，在此区域内溶菌酶溶液分为两种液相。他们利用光散射技术为这种分离制作了相图。为了估计忽略对流的溶液层厚度，a - group对晶体生长时溶液层厚度变化的浓度分布进行了计算机模拟。j组测量了生长中的溶菌酶晶体周围的浓度分布，改变了细胞的厚度。这些结果相互比较。j组测定HEW溶菌酶在高压下的原位溶解度。还研究了压力对生长速率的依赖性。少

Achievements of this year are as follows ;1. The Japanese group (J-group) measured solubilities of tetragonal, orthorhombic and triclinic form of hen egg white lysozyme (HEW lysozyme) using two beam interferometry.2. J-group developed an interferometic method to measure the refractive index inside crystals (HEW lysozyme).3. J-group got highly purified HEW lysozyme samples using HPLC.With this sample, they observed crystal interfaces using atomic force microscopy. This result was discussed with the American group (A-group) comparing with A-group's result.4. J-group tried to perform crystallization of HEW lysozyme in a high magnetic field. And they found that it could control the nucleation rate and orientation of crystals.5. Appoferritin (A-group) and ferritin (J-group) were highly purified and then crystallized. They showed again that removement of impurities is indispensable for growth of crystals of high quality. J-group made a morphodrom of ferritin.6. A-group performed light scatte … More ring experiments of HEW lysozyme solution and discuss the aggregation process. They estimated the second virial coefficient of the solution and discussed relation between ion intensity and intermolecular interaction.7. A-group precisely measured topography of vicinal surface with use of the laser interferometry and obtained time evolution of normal and step growth rates and slope of the vicinal surfaces.8. A-group found that there existed a region where HEW lysozyme solution separated into two liquid phases. They made a phase diagram for this segregation with use of the light scattering technique.9. In order to estimate the solution layr thickness where convection is ignored, A-grup performed computer simulation of the concentration profiles around a growing crystal changing thickness of the solution layr. J-group measured concentration profiles around a growing lysozyme crystal changing the thickness of the cell. These results were compared each other.10. J-group measured solubility of HEW lysozyme in situ in high pressure. Pressure dependence of the growth rate was also studied. Less