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

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.

在本研究中，利用原位原子力显微镜 (AFM) 和荧光显微镜 (FM) 研究了杂质对蛋白质（鸡蛋清溶菌酶）晶体生长的影响。主要结果如下： 1． AFM观察溶液交换系统的开发。我们先后开发了利用AFM观察蛋白质分子吸附过程的溶液交换系统。2．蛋白质在晶体表面吸附过程的观察。利用上述系统直接观察了（纯）溶菌酶在晶体表面的吸附情况。 3．蛋白质杂质引起的蛋白质晶体的形态变化我们通过AFM观察了被卵转铁蛋白污染的四方溶菌酶晶体的表面，卵转铁蛋白的分子量比溶菌酶的分子量大得多，并发现卵转铁蛋白优先吸附在（101）上。 FM观察还表明，(101)表面附近的杂质密度相对高于其他区域。这些结果解释了溶菌酶晶体引起的蛋白质晶体的形态变化。

项目成果

内山幸昌, 水谷武臣, 中田俊隆: "原子間力顕微鏡によるタンパク質吸着過程のその場観察"立命館大学理工学研究所紀要. 60. (2001)

Yukimasa Uchiyama、Takeomi Mizutani、Toshitaka Nakata：“使用原子力显微镜对蛋白质吸附过程进行原位观察”立命馆大学科学技术研究所通报60。（2001）。

内山幸昌, 水谷武臣, 中田俊隆: "原子間力顕微鏡によるタンパク質吸着過程のその場観察"立命館大学理工学研究所紀要. 61. (2002)

Yukimasa Uchiyama、Takeomi Mizutani、Toshitaka Nakata：“使用原子力显微镜对蛋白质吸附过程进行原位观察”立命馆大学科学技术研究所通报61。（2002）。

Y. Uchiyama, T. Mizutani and T. Nakada: "In-situ observation of protein adsorption process by atomic force microscopy"Memories of the institute of Science & engineering Ritsumeikan Univ. Shiga, Japan. 60. (2001)

Y. Uchiyama、T. Mizutani 和 T. Nakada：“用原子力显微镜原位观察蛋白质吸附过程”科学研究所的回忆