Structure and function of low-molecular-weight proteins with a cytochrome P450 heme-binding domain derived from the basidiomycete Lentinus edodes

担子菌香菇来源的具有细胞色素 P450 血红素结合域的低分子量蛋白的结构和功能

• 批准号: 10660076
• 负责人: SHISHIDO Kazuo
• 金额: $ 2.05万
• 依托单位: Tokyo Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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-10660076/
• 关键词: basidiomycete; cytochrome P450; heme-binding domain; repeat sequence; low-molecular-weight heme protein; チトクロムP450; 一酸化炭素差スペクトル

A novel gene shp1 isolated from the basidiomycete Lentinus edodes DDLE1 encodes a low-molecular-weight protein with a cytochrome P450 heme-binding domain. The 5' untranslated region of shp1 contained a repeat sequence (named LESR1) highly interspersed over L. edodes genome, which is transcribed together with the coding region as a single transcript. The SHP1 protein consisted of 74 amino acids residues and contained the region highly homologous to the heme-binding regions of L. edodes P450s (500 amino acids) isolated from FMC2 strain in this study. The shp1 isolated from L. edodes FMC2 was found to encode 115 amino acids. The DDLE1 SHP1 protein produced in E. coli was mixed with protoheme in the presence of deoxycholic acid, followed by the treatment of reconstitution of the protein-protoheme complex. Carbon monooxide difference spectrum of the mixture suggests that the SHP1 is a heme-binding protein with a similarity to P450s in the type of heme binding. Then we attempted to determine the substrate and reaction product of SHP1 by using S. cerevisiae system used for the detection of Aflatoxin B1 generating activity of the ascomycetous fungal P450s. Unfortunately the substrate for SHP1 hasn't been determined until now.

一个新的基因shp 1分离自担子菌香菇DDLE 1编码一个低分子量蛋白与细胞色素P450血红素结合域。shp 1的5'端非翻译区含有一个高度散布在L. edodes基因组，其与编码区一起转录为单个转录物。SHP 1蛋白由74个氨基酸残基组成，含有与L. edodes P450（500个氨基酸）。从L.香菇FMC 2基因编码115个氨基酸。在E.在脱氧胆酸存在下，将大肠杆菌与血红素混合，然后进行蛋白质-血红素复合物的重构处理。混合物的一氧化碳差谱表明，SHP 1是一种血红素结合蛋白，与P450的血红素结合类型相似。然后尝试用S.酿酒酵母系统用于检测子囊真菌P450的黄曲霉毒素B1产生活性。不幸的是，SHP 1的底物直到现在还没有确定。

项目成果

秋山涼子: "担子菌シイタケのシトクロムP450及び関連ヘム蛋白質遺伝子のクローニングと解析"東京工業大学大学院バイオサイエンス専攻修士課程要旨集. 41-42 (2000)

Ryoko Akiyama：“担子菌香菇中细胞色素P450和相关血红素蛋白基因的克隆和分析”东京工业大学生物科学研究生院硕士课程摘要41-42（2000）。

秋山涼子、佐藤義弘、梶原将、宍戸和夫: "担子菌シイタケからの2つのcytochromeP450遺伝子の単離と解析"第22回日本分子生物学会年会講演要旨集. -538 (1999)

Ryoko Akiyama、Yoshihiro Sato、Masashi Kajiwara、Kazuo Shishido：“来自担子菌香菇的两个细胞色素 P450 基因的分离和分析”日本分子生物学会第 22 届年会记录 -538（1999 年）。

秋山涼子: "担子菌シイタケのシトクロムP450及び関連ヘム蛋白質遺伝子のクローニングと解析"東京工業大学大学院バイオサイエンス専攻修士論文要旨集. 41-42 (2000)

Ryoko Akiyama：“担子菌香菇细胞色素 P450 和相关血红素蛋白基因的克隆和分析”东京工业大学生物科学系硕士论文摘要 41-42 (2000)。

Ryouko AKIYAMA, Yoshihito SATO, Susumu KAJIWARA, and Kazuo SHISHIDO: "Isolation and characterization of two genes encoding cytochrome P450s from the basidiomycete Lentinus edodes"Abstract of the 22nd Annual Meeting of Japanese Society of Molecular Biologi

Ryouko AKIYAMA、Yoshihito SATO、Susumu KAJIWARA 和 Kazuo SHISHIDO：“从担子菌香菇中编码细胞色素 P450 的两个基因的分离和表征”日本分子生物学学会第 22 届年会摘要

Ryouko AKIYAMA: "Isolation and characterization of the basidiomycete Lentinus edodes genes encoding cytochrome P450s and related heme proteins"Abstract of the Presentations for Master Degree in Department of Life Science, Graduate School of Bioscience and

Ryouko AKIYAMA：“担子菌香菇基因编码细胞色素 P450 和相关血红素蛋白的分离和特性”生物科学研究生院生命科学系硕士学位论文摘要