Nucleation and Growth Kinetics of Protein Crystals

蛋白质晶体的成核和生长动力学

• 批准号: 09044055
• 负责人: KOMATSU Hiroshi
• 金额: $ 2.11万
• 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: 1997
• 资助国家: 日本
• 起止时间: 1997 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09044055/
• 关键词: Crystal Growh; Protein; Hen Egg-White Lysozyme; Two-beam Interferometry; Solubility; Micro-gravity; Magnetic Field Effects; Impurity Effects

Results obtained by this project were summarized as below.(1) Japanese team developed a Michelson Interferometer objective lens of super-long working distance (25.5 mm) with Nikon Co. Using this interferometer, a novel technique for determining solubility was developed, and solubilities of tetragonal and orthorhombic lysozyme crystal were detemined as a function of temperature for the first time.(2) Japanese team studied magnetic field effects on the crystallization of lysozyme, and found that a magnetic field (i) increased a growth rate and (ii) decreased a solubility. Equipment to measure magnetic birefringence was developed.(3) Japanese team highly-purified commercial lysozyme (99.9%) by purification technique developed by U.S.A.team. Growthprocess of lysozyme crystals were in situ observed by atomic force microscopy. It was found that small amount of impurity (1%) drastically decreased growth rate of (101) face of tetragonal crystal.(4) Japanese team developed a novel technique to measure impurity content and its distribution in the crystal with a fluorescense indicator. A novel mechanism of the impurity incorporation into the crystal was proposed.(5) U.S.A.team analyzed the concentration distribution around growing lysozyme crystals using computer simulation technique. Japanese team measured the actual concentration distribution around the crystal by interferometry. These two results were compared each other and found that they mostly agreed. It was found that buoyancy driven convection can be stopped when the thickness of the solution is smaller than l50オm.(6) Japanese team observed the concentration distribution around lysozyme crystals under lg to micro-g using free fall facilities. The changes in the concentration distribution were analyzed.

本项目取得的成果总结如下：（1）日本团队与尼康公司合作开发了超长工作距离（25.5mm）的迈克尔逊干涉仪物镜。利用该干涉仪，开发了一种测定溶解度的新技术，并研究了溶解度的测定方法。首次测定了四方晶系和斜方晶系溶菌酶晶体的结晶度随温度的变化。(2)日本团队研究了磁场对溶菌酶结晶的影响。溶菌酶，并发现磁场（i）增加了生长速率，（ii）降低了溶解度。开发了测量磁双折射的设备。（3）日本团队通过开发的纯化技术高度纯化了商业溶菌酶（99.9％）。美国团队通过原子力显微镜原位观察溶菌酶晶体的生长过程，发现少量杂质（1%）会显着降低溶菌酶晶体的生长速度。四方晶体的（101）面。（4）日本团队开发了一种利用荧光指示剂测量晶体中杂质含量及其分布的新技术。提出了杂质掺入晶体的新机制。（5）美国团队。日本团队利用计算机模拟技术分析了生长的溶菌酶晶体周围的浓度分布，通过干涉测量法测量了晶体周围的实际浓度分布，发现它们基本一致。发现当溶液厚度小于l50欧姆时，浮力驱动的对流可以停止。(6)日本研究小组利用自由落体装置观察了1g至微g下溶菌酶晶体周围的浓度分布，并观察了浓度分布的变化。分析了。

项目成果

B.R.Thomas et al.: "Heterogeneity determination and purification of commercial hen egg white lysozyme" Acta Cryst.D52. 776-784 (1996)

B.R.Thomas 等人：“商业鸡蛋清溶菌酶的异质性测定和纯化”Acta Cryst.D52。

B.R.Thomas et al.: "Heterogeneity determinationand purification of commercial hen egg white lysozyme" Acta Crystalography. D52. 776-784 (1996)

B.R.Thomas 等人：“商品鸡蛋清溶菌酶的异质性测定和纯化”Acta Crystalography。

F.Rosenberger et al.: "A rationale forsystem-dependent advantages and disadvantages of solution crystal growth atlow gravity" Microgravity Sci.Technol.(印刷中).

F. Rosenberger 等人：“低重力下溶液晶体生长的系统相关优点和缺点的基本原理”微重力科学（正在出版）。

H.Komatsu et al.: "“An atomic level analysis of crystal growth mechanism in complex systems by means of nano-optical microscopy and AFM"in Advances in crystal growth mechanisms" Elsevier,Amsterdam, 15 (1997)

H. Komatsu 等人：“晶体生长机制进展中的“通过纳米光学显微镜和 AFM 对复杂系统中晶体生长机制的原子水平分析””Elsevier，阿姆斯特丹，15 (1997)

K.Kurihara et al.: "Interferometric study on crystal growth of tetragonal lysozyme crystal" Journal of Crystal Growth. 166. 904-908 (1996)

K.Kurihara 等人：“四方溶菌酶晶体晶体生长的干涉研究”《晶体生长杂志》。