物理化学学报 >> 2001, Vol. 17 >> Issue (06): 531-536.doi: 10.3866/PKU.WHXB20010612

研究论文 上一篇    下一篇

气相扩散法生长溶菌酶晶体的动态光散射研究

戴国亮;董向群;孙祉伟;胡文瑞   

  1. 中国科学院力学研究所 国家微重力实验室,北京 100080
  • 收稿日期:2000-11-17 修回日期:2001-03-02 发布日期:2001-06-15
  • 通讯作者: 戴国亮 E-mail:dspr@imech.ac.cn

Study on Lysozyme Crystal Growth by Vapor Diffusion Approach Using Dynamic Light Scattering Method

Dai Guo-Liang;Dong Xiang-Qun;Sun Zhi-Wei;Hu Wen-Rui   

  1. National Microgravity Laboratory, Institute of Mechanics, Chinese Academy of Sciences, Beijing 100080
  • Received:2000-11-17 Revised:2001-03-02 Published:2001-06-15
  • Contact: Dai Guo-Liang E-mail:dspr@imech.ac.cn

摘要: 首次采用动态光散射研究了气相扩散法生长溶菌酶晶体.实验中采用了两种溶解溶菌酶的方法,所得实验结果是有区别的.这种区别表明了NaCl对溶菌酶分子间相互作用产生十分重要的影响.实验结果表明,晶体生长过程中,溶液中溶菌酶始终保持单分子与两分子聚集体的状态,这种状态是生长晶体的基础.

关键词: 动态光散射, 气相扩散法, 晶体生长, 溶菌酶, 扩散系数, 批量法

Abstract: Both vapor diffusion approach and batch mode are often used approaches for protein crystallization. The light scattering method is a powerful tool particularly for prenucleation study. However, all light scattering studies were limited to the crystallization process of the batch mode. To apply dynamic light scattering technique to the protein crystallization study for the vapor diffusion approach, as well as the batch mode, a special sample cell (see Fig.1) was desired in the present work. It has two separate parts, a scattering cell and a thin tube filled with salt solution. The results showed that our special cell was reliable for light scattering study(see table 1). In our experiments, vapor diffusion approach was mainly used. Sodium acetate buffer solution, 0.04 mol•L-1, pH=4.5, was mixed with sodium chloride,whose final concentration was 0.345 mol•L-1. Chicken egg white lysozyme was dissolved in the above solution with a concentration of 50 mg•mL-1 and 25 mg•mL-1. Two lysozyme dissolution ways were used. One was lysozyme dissolved in only sodium acetate buffer solution and another is in buffer solution with sodium chloride. Their results were different(see Fig.2 and Fig.5), that showed NaCl played an important role in the interaction of lysozyme molecules. Due to the feature of vapor diffusion approach, it was verified that the force between lysozyme moleculars in the solution is weak during the whole crystallization process, which is desirable for crystal growth, namely, lysozyme molecules kept in monomolecule or aggregation of bimolecules in solution when lysozyme crystal grew. Besides, our results show that there is a significant difference between this approach and the batch mode(decreasing temperature) even if other parameters are the same. This significant difference is the direct result of different ways of supersaturation. Vapor diffusion approach, compared to batch mode is a more temperate, precisely controlled way to realize supersaturation.

Key words: Dynamic light scatter, Vapor diffusion approach, Crystal growth, Lysozyme, Diffusion coefficient, Batch method