Growth of High Quality Protein Crystals using Magnetic Field

利用磁场生长高质量蛋白质晶体

• 批准号: 10555001
• 负责人: SAZAKI Gen
• 金额: $ 8万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10555001/
• 关键词: protein; crystal; magnetic field; orientation; lysozyme; crystal growth; convection; crystal perfection; protein; crystal; magnetic field; orientation; lysozyme; crystal growth; convection; crystal perfection; oriontation

We chose hen egg-white lysozyme as a model protein, and studied a magnetic field effects on the crystallization of lysozyme crystals. Main results obtained are summarized as follows.1. Magnetic orientation of the crystalsMagnetic orientation of protein crystals could be controlled by changing the growth rate and depth of the crystallization container in addition to the magnetic field strength.2. Growth rate of the crystalsIn situ observation of growing protein crystals under 11T revealed that a magnetic field decreased the growth rate of the crystals.3. Damping of convection in NaCl aqueous solutionIn situ observation of temperature-driven convection in NaCl aqueous solution showed that a magnetic field damped the convection even in the case of "aqueous solution".4. Improvement of the quality of the crystalRocking curve measurement of orthorhombic lysozyme crystals grown under 10T revealed that a magnetic field drastically increased the "crystal perfection" of protein single crystal., and gave higher diffraction limit and smaller X-ray radiation damage.5. Effects of other external force I : high pressureWith an increase in hydrostatic pressure, solubility of tetragonal lysozyme crystals increased, though that of orthorhombic one decreased. Increase in surface energy decreased the growth rate of tetragonal and orthorhombic crystals.6. Effects of other external force II : microgravityMicrogravity experiments using dropping facility revealed that 1) Grashoff number could be a good measure to predict buoyancy convection, and that 2) Marangoni convection at water-air interface decreased to less than 10% within 10-15 min because of the contamination of the interface.

以蛋清溶菌酶为模型蛋白，研究了磁场对溶菌酶晶体结晶的影响。主要研究成果总结如下：1.研究成果。晶体的磁取向除了磁场强度外，还可以通过改变结晶容器的生长速度和深度来控制蛋白质晶体的磁取向。晶体生长速度在11T下对生长的蛋白质晶体进行原位观察发现，磁场降低了晶体的生长速度。氯化钠水溶液中对流的衰减对温度驱动的水溶液中对流的现场观测表明，即使在“水溶液”的情况下，磁场也对对流有抑制作用。对10T以下生长的正交型溶菌酶晶体摇摆曲线测量质量的改进表明，磁场显著提高了蛋白质单晶的晶体完整性，并给出了更高的衍射极限和更小的X射线辐射损伤。其他外力的影响I：高压随着静水压力的增加，四方溶菌酶晶体的溶解度增加，而正交溶菌酶晶体的溶解度降低。表面能的增加降低了四方和斜方晶体的生长速度。微重力实验表明：1)Grashoff数可以很好地预测浮力对流；2)由于界面的污染，水-气界面的Marangoni对流在10-15分钟内下降到10%以下。

项目成果

Y.Suzuki,et al.: "Interferometric study of the solubility of lysozyme crystal under high pressure"Journal of Crystal Growth. 209. 1018-1022 (2000)

Y.Suzuki等人：“高压下溶菌酶晶体溶解度的干涉研究”晶体生长杂志。

G.Sazaki et al.: "Solubility of tetragonal and orthorhombic lysozyme crystals under high pressure"Journal of Crystal Growth. 196. 204-209 (1999)

G.Sazaki等人：“高压下四方晶系和斜方晶系溶菌酶晶体的溶解度”晶体生长杂志。

Y. Suzuki, T. Sawada, S. Miyashita, H. Komatsu, G. Sazaki, T. Nakada: "Interferometric study of the solubility of lysozyme crystal under high pressure"J. Crystal Growth. 209. 1018-1022 (2000)

Y. Suzuki、T. Sawada、S. Miyashita、H. Komatsu、G. Sazaki、T. Nakada：“高压下溶菌酶晶体溶解度的干涉研究”J。

G.Sazaki et al.: "Magnetic damping of the temperature-driven convection in NaCl aqueous solution using a static and homogeneous field of 1OT"Japanese Journal of Applied Physics. 38. L842-L844 (1999)

G.Sazaki 等人：“使用 1OT 静态均匀场对 NaCl 水溶液中温度驱动对流进行磁阻尼”《日本应用物理学杂志》。

G.Sazaki,et al: "Solubility of tetragonal and orthorhombic lysozyme crystals under high pressure" Journal of Crystal Growth. 196. 204-209 (1999)

G.Sazaki 等人：“高压下四方晶系和斜方晶系溶菌酶晶体的溶解度”《晶体生长杂志》。