Impurity effects of protein oligomer for protein crystallization and crystal quality

蛋白质低聚物的杂质对蛋白质结晶和晶体质量的影响

• 批准号: 11650023
• 负责人: NAKADA Toshitaka
• 金额: $ 0.83万
• 依托单位: Ritsumeikan University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11650023/
• 关键词: Atomic force microscopy; Protein; Growth step; Fluorescent; Lysozyme; Impurity; crystal; Atomic force microscopy; Protein; Growth step; Fluorescent; Lysozyme; Impurity; crystal

In this study, impurity effects on the growth of protein (hen egg white lysozyme) crystals have been studied using in-situ atomic force microscopy (AFM) and fluorescent microscopy (FM). The main results are as follows;1. Development of solution exchange system for AFM observations.We successively developed a solution exchange system to observe adsorption process of protein molecules by using AFM.2. Observation of adsorption process of protein on crystal surfaces.The adsorption of (pure) lysozyme to crystal surfaces has been directly observed by using the system described above.3. Morphology changes of protein crystals induced by a protein impurityWe observed surfaces of tetragonal lysozyme crystals contaminated by ovotransferrin, which has rather larger molecular weight than that of lysozyme by AFM, and revealed that ovotransferrin preferentially adsorbs on the (101). FM observation also showed that the density of the impurities near the (101) surface is relatively higher than the other region. These results explain the morphological changes of protein crystals induced by the lysozyme crystals.

在这项研究中，使用原子原子力显微镜（AFM）和荧光显微镜（FM）研究了对蛋白质（Hen Egg White溶菌酶）晶体生长的杂质影响。主要结果如下; 1。为AFM观测的开发解决方案交换系统。我们依次开发了一种解决方案交换系统，以使用AFM.2观察蛋白质分子的吸附过程。2。通过使用上述系统直接观察到（纯）溶菌酶对晶体表面的吸附过程。3。由蛋白质杂质诱导的蛋白质晶体的形态变化观察到了被卵晶氧蛋白污染的四方溶菌酶晶体的表面，该蛋白晶体的分子量比AFM的分子量更大，并揭示了Ovotransfransfrins fysozyme的形态。 FM观察结果还表明，（101）表面附近的杂质的密度相对较高。这些结果解释了溶菌酶晶体诱导的蛋白质晶体的形态变化。