X-Ray Crystallographic and Computer Graphic Analyses of the Mammalian Cytochrome P-450's and Their Mutants

哺乳动物细胞色素 P-450 及其突变体的 X 射线晶体学和计算机图形分析

• 批准号: 01044016
• 负责人: HATANO Masahiro
• 金额: $ 2.3万
• 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: 1989
• 资助国家: 日本
• 起止时间: 1989 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01044016/
• 关键词: Cytochrome P-450; X-ray crystallography; Site-directed mutagenesis; Computer graphics; Fuction-structure relationship; Heme-protein; Amino acid substitutions; Recombinant DNA

A family of protoheme-containing enzymes, collectively called cytochrome P-450(P-450), are involved in a variety of oxidative reactions. Eukaryotic P-450 is a membranebound enzyme, and thus its tertiary structure may differ somewhat from the crystal structure of the soluble bacterial P-450_<cam> decided by Poulos. We had previously developed an efficient expression system for rat liver P-450_d in the Saccharomyces cerevisiae. With site-directed mutageneses, the roles of each amino acids were clarified.

含有原血红素的酶家族，统称为细胞色素 P-450（P-450），参与多种氧化反应。真核生物P-450是一种膜结合酶，因此其三级结构可能与Poulos确定的可溶性细菌P-450_<cam>的晶体结构有所不同。我们之前在酿酒酵母中开发了大鼠肝脏 P-450_d 的高效表达系统。通过定点诱变，每种氨基酸的作用都得到了阐明。

项目成果

旗野昌弘(分担執筆): "タンパク質工学" 丸善株式会社, 211 (1989)

波多野正宏（撰稿人）：《蛋白质工程》丸善株式会社，211（1989）

H.Furuya,T.Shimizu,K.Hirano,M.Hatano,Y.Fujii-Kuriya,R.Raag,and T.L.Poulos: "Site-Directed Mutageneses of Rat Liver Cytochrome P-450_d:Catalytic Activities toward Benzphetamine and 7-Ethoxy-Coumarin" Biochemistry. 28. 6848-6857 (1989)

H.Furuya、T.Shimizu、K.Hirano、M.Hatano、Y.Fujii-Kuriya、R.Raag 和 T.L.Poulos：“大鼠肝脏细胞色素 P-450_d 的定点诱变：对苯异丙胺和 7- 的催化活性

Abu Jafar Md.Sadeque,T.Shimizu,K.Hirano,M.Hatano,and Y.Fujii-Kuriyama: "Interaction of Fluoroaniline with Cytochrome P-450_d Mutants:Difference Absorption Spectral Studies" Inorganica Chmica Acta. 153. 161-164 (1988)

Abu Jafar Md.Sadeque、T.Shimizu、K.Hirano、M.Hatano 和 Y.Fujii-Kuriyama：“氟苯胺与细胞色素 P-450_d 突变体的相互作用：差异吸收光谱研究”无机化学学报。

H.Furuya,T.Shimizu,M.Hatano,Y.Fujii-Kuriyama: "Mutations at the Distal and Proximal Sites of Cytochrome P-450_d Changed Regio-Specificity of Acetanilide Hydroxylations" Biochemical and Biophysical Research Communications. 160. 669-676 (1989)

H.Furuya，T.Shimizu，M.Hatano，Y.Fujii-Kuriyama：“细胞色素 P-450_d 远端和近端位点的突变改变了乙酰苯胺羟基化的区域特异性”生物化学和生物物理研究通讯。

Hideo Sotokawa, Toru Shimizu, Hideyuki Furuya, Abu Jafar Md. Sadeque, Masahiro Hatano, Yasunori Ohba, Masamoto Iwaizumi, and Yoshiaki Fujii-Kuriyama: "Electron Spin Resonance Studies of Wild-Type and Mutant Cytochromes P-450_d: Effects of Mutations at Pro

Hideo Sotokawa、Toru Shimizu、Hideyuki Furuya、Abu Jafar Md. Sadeque、Masahiro Hatano、Yasunori Ohba、Masamoto Iwaizumi 和 Yoshiaki Fujii-Kuriyama：“野生型和突变型细胞色素 P-450_d 的电子自旋共振研究：突变的影响