In situ observation of protein crystallization under high pressure

高压下蛋白质结晶的原位观察

• 批准号: 04452321
• 负责人: KOMATSU Hiroshi
• 金额: $ 3.78万
• 依托单位: Institution for Material Research, Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04452321/
• 关键词: HEW lysozyme; a high pressure cell; nucleation control by pressure; habit change by pressure; solubility; crystal growth of biological macromolecules; リゾチーム; タンパク質に及ぼす高圧効果; 高圧溶液中の結晶成長; ダイヤモンドアンビル; 濃度分布の測定; リゾチームと多形

1. Designing and build-in of the high pressure in situ observation system :A diamond anvil cell was designed to fit the metallurgical phase-contrast microscope. The pressure measurements system was built up using the fluorescence shift of a spectrum from a ruby laser beam.2. A possible way of controlling the number of nuclei by applying pressure on HEW lysozyme crystals was found.It was found that the number of nuclei of HEW lysozyme diminishes when the nucleation took place under pressure. The higher the pressure the less number of nuclei appeared.3. The pressure dependence of the solubility of HEW lysozyme :The pressure was changed from 510MPa to 0.1 MPa at 25(〕SY.+-.〔)0.2ﾟC using the anvil. It was found that the single crystals of HEW lysozyme (tetragonal system) grew as the pressure was decreased. It became clear for the first time that the solubility of HEW lysozyme increased when the pressure in the vessel was increased.4. Habit change of HEW lysozyme by applying pressure :The ratio of growth rate vertical to the faces (101) and (110) increased monotonously as the pressure was increased. This resulted in the habit change of the crystal ; the bulk crystals c axis.

1. 高压原位观测系统的设计与搭建：为配合冶金相衬显微镜，设计了金刚石顶砧单元。利用红宝石激光束的光谱荧光位移建立了压力测量系统。发现了一种通过对HEW溶菌酶晶体施加压力来控制细胞核数量的可能方法。在压力下成核时，HEW溶菌酶的细胞核数量减少。压力越大，出现的核数越少。HEW溶菌酶溶解度的压力依赖性：在25(]SY.+-.[)0.2‰的温度下，用顶砧将压力从510MPa改变为0.1 MPa。结果表明，随着压力的降低，HEW溶菌酶（四边形体系）的单晶逐渐增多。首次发现HEW溶菌酶的溶解度随着容器内压力的增加而增加。施加压力对HEW溶菌酶习性的影响：随着压力的增加，垂直于表面的生长率（101）和（110）的比值单调增加。这导致晶体的习惯改变；体晶体c轴。

项目成果

鈴木 良尚: "高圧下でのHEWリゾチームの結晶成長" 日本結晶成長学会誌. 20. 168-168 (1993)

Yoshihisa Suzuki：“高压下 HEW 溶菌酶的晶体生长”日本晶体生长学会杂志 20. 168-168 (1993)。

Satoru Miyashita, Hiroshi Komatsu, Yoshihisa Suzuki and Toshitaka Nakada: "Observation of the concentration distribution around a growing lysozyme crystal." J.Crystal Growth. to be published. (1994)

Satoru Miyashita、Hiroshi Komatsu、Yoshihisa Suzuki 和 Toshitaka Nakada：“观察正在生长的溶菌酶晶体周围的浓度分布。”

Hiroshi Komatsu: "Interferometric Observation of the Interfacial Gradient Layers Around a Lysozyme Crystal." Jpn.J.Appl.Phys.32. L1855-L1857 (1993)

Hiroshi Komatsu：“溶菌酶晶体周围界面梯度层的干涉观测”。

Hiroshi Komatsu, Satoru Miyashita and Yoshihisa Suzuki: "Interferometric observation of the interfacial gradient layrs around a lysozyme crystal." Jpn. J.Appl. Phys.32. L1855-L1857 (1993)

Hiroshi Komatsu、Satoru Miyashita 和 Yoshihisa Suzuki：“溶菌酶晶体周围界面梯度层的干涉观测。”

Satoru Miyashita, Hiroshi Komatsu, Yoshihisa Suzuki and Toshitaka Nakada: "Observation of the concentration distribution around a growing lysozyme crystal." Proc. 7th Topical Meeting on Cryst. Growth. Mech.407-412 (1994)

Satoru Miyashita、Hiroshi Komatsu、Yoshihisa Suzuki 和 Toshitaka Nakada：“观察正在生长的溶菌酶晶体周围的浓度分布。”