Nucleation and Growth Kinetics in Protein Crystallization

蛋白质结晶中的成核和生长动力学

• 批准号: 07044057
• 负责人: KOMATSU Hiroshi
• 金额: $ 5.89万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07044057/
• 关键词: proteins; lysozyme; crystal growth; impurity effect; two beam interferometry; light scattering; solubility; effect of magnetic field; タンパク質; 濃度分布

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

今年取得的成果如下： 1．日本课题组（J组）利用两束干涉法测定了鸡蛋清溶菌酶（HEW溶菌酶）的四方晶系、斜方晶系和三斜晶系的溶解度。2．测量晶体内部的折射率（HEW溶菌酶）。3．利用该样品，他们使用原子力显微镜观察了晶体界面，并与美国组（A组）讨论了该结果，并与A组的结果进行了比较。4．他们发现它可以高度控制晶体的成核率和取向。5．他们再次表明，去除杂质对于制备高质量铁蛋白晶体是必不可少的。6．他们估计了溶液的第二维里系数并讨论了离子强度与分子间相互作用之间的关系。7． 8．A组发现存在一个HEW溶菌酶溶液分离成两个液相的区域。使用光散射技术进行这种分离。9. 为了估计忽略对流的溶液层厚度，A 组对 J 组的生长晶体变化厚度周围的浓度分布进行了计算机模拟。测量的生长的溶菌酶晶体周围的轮廓改变了细胞的厚度。J组还研究了高压下原位溶解度的测量。

项目成果

T.Nakada, G.Sazaki, S.Miyashita and H.Komatsu: "In situ observation of growing surface of lysozyme crystal" Journal of Crystal Growth. (In preparation).

T.Nakada、G.Sazaki、S.Miyashita 和 H.Komatsu：“溶菌酶晶体生长表面的原位观察”《晶体生长杂志》。

F.Rosenberger,P.G.Vekilov,M.Muschol and B.R.Thomas: "Nucleation and crystallization of globular proteins-what do we know and what is missing?" J.Crystal Growth. 168. 1-27 (1996)

F.Rosenberger、P.G.Vekilov、M.Muschol 和 B.R.Thomas：“球状蛋白的成核和结晶——我们知道什么以及缺少什么？”

G.Sazaki, E.Yoshida, H.Komatsu, T.Nakada, S.Miyashita and K.Watanabe: "Effects of a magnetic field on the nucleation and growth of protein crystals" Journal of Crystal Growth. (in press).

G.Sazaki、E.Yoshida、H.Komatsu、T.Nakada、S.Miyashita 和 K.Watanabe：“磁场对蛋白质晶体成核和生长的影响”《晶体生长杂志》。

M.Muschol and F.Rosenberger: "Lack of evidence for prenucleation aggregate formation in lysozyme crystal growth solutions." J.Crystal Growth. 167. 738-747 (1996)

M.Muschol 和 F.Rosenberger：“缺乏溶菌酶晶体生长溶液中成核前聚集体形成的证据。”

G.Sazaki,E.Yoshida,H.Komatsu,T.Nakada,S.Miyashita and K.Watanabe: "Effects of a magnetic field on the nucleation and growth of protein crystals" Journal of Crystal Growth. (印刷中).

G. Sazaki、E. Yoshida、H. Komatsu、T. Nakada、S. Miyashita 和 K. Watanabe：“磁场对蛋白质晶体成核和生长的影响”《晶体生长杂志》（出版中）。