Elemental Analysis and Crystallization of Protein Crystals

蛋白质晶体的元素分析和结晶

• 批准号: 01490017
• 负责人: SATO Kiyotaka
• 金额: $ 3.71万
• 依托单位: Hiroshima 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: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-01490017/
• 关键词: Protein crystallization; Lattice defect; Toxic protein; Algae lectin; Secreted protein; タンパク質晶化; 結晶欠陥; 毒性タンパク質; たんぱく質; 結晶化; 植物細胞分泌タンパク質; 卵白リゾチ-ム; 光散乱トモグラフ法; 有用たんぱく質; たんぱく質の結晶化; lepidopteran toxin; Ado Hcy hydrolase; メチル基転移反応酵素

Crystallization processes of a model substance : hen-egg white lysozyme were analyzed with laser scattering tomography, etching technique, heterogeneous crystallization and optical microscopy. The crystallization kinetics of an orthorhombic modification was linearly dependent in a logarithmic manner on the concentration of precipitants. The effects of polymorphs were observed in the crystallization kinetics. Lattice defects in lysozyme crystals were examined with laser scattering tomography and etching technique. The defects of point type were concentratedly distributed closer to the growing surfaces and growth sectors were observed. Heterogeneous crystallization on glass and polymer substrates was tried to selectively grow single crystals.Crystallization of three proteins was newly challenged : toxin protein yielded in bacteria, vegetable cell-producing protein and lectin produced from algae. Pure samples of about 1 0 mg yield of each substance was obtained from raw materials, and subjected to crystallization using vapor diffusion method. However, amorphous solids have so far been obtained. Further trials are needed for these substances.

采用激光散射层析、蚀刻技术、非均相结晶和光学显微镜等方法对鸡蛋白色溶菌酶的结晶过程进行了研究。斜方晶改性的结晶动力学是线性依赖于在对数的方式上的沉淀剂的浓度。在结晶动力学中观察到多晶型的影响。用激光散射层析术和腐蚀技术研究了溶菌酶晶体中的晶格缺陷。点型缺陷集中分布在靠近生长面处，并观察到生长扇区。在玻璃和聚合物衬底上的非均相结晶尝试了选择性生长单晶，其中细菌产生的毒素蛋白、植物细胞产生的蛋白和藻类产生的凝集素是三种蛋白质结晶的新挑战。从原料中获得每种物质约10 mg产率的纯样品，并使用气相扩散法进行结晶。然而，迄今为止已经获得了无定形固体。需要对这些物质进行进一步的试验。

项目成果

浜中 完範,佐藤 清隆: "卵白リゾチ-ム結晶化の速度論的解析" 日本結晶成長学会誌. 17. 110-115 (1990)

Mitsunori Hamanaka、Kiyotaka Sato：“蛋清溶菌酶结晶的动力学分析”日本晶体生长学会杂志 17. 110-115 (1990)。

K.Hori,R.Okamoto,K.Miyazawa,K.Ito: "Isolation and characterization of a new hemaglutimen from the red alga <Gracilaria bursaーpactoriz>___ー" Experientia. 46. 975-976 (1990)

K.Hori、R.Okamoto、K.Miyazawa、K.Ito：“从红藻中分离和表征新的血凝素 <Gracilaria bursapactoriz>___”Experientia。 46. 975-976 (1990)

M.Esaka: "Molecular cloning and nucliotide Sequence of fullーlength cDNA for ascorbate oxidase from culturea pumpkin cells" Eur.J.Biochem.191. 537-541 (1990)

M.Esaka：“来自培养南瓜细胞的抗坏血酸氧化酶的全长 cDNA 的分子克隆和核苷酸序列”Eur.J.Biochem.191 (1990)。

K.Sato,Y.Fukuba,T.Mitsuda et al.: "Observation of Lattice defects in Orthorbombic Hen.Egg White Lysozyme Crystals with Laser Scattering" J.Crystal Growth.

K.Sato、Y.Fukuba、T.Mitsuda 等人：“用激光散射观察正交母鸡蛋清溶菌酶晶体中的晶格缺陷”J.Crystal Growth。

Y. Fukuba & K. Sato: "Heterogeneous nucleation and single crystal growth of hen egg-white lysozyme on glass plates" J. Fac. Appl. Biol. Sci. Hiroshima Univ.30. 129-134 (1991)

Y·福波