The Function of B Cytochrome in Complex II (Succinate-Ubiquinone Oxidoreductase)

复合物 II（琥珀酸泛醌氧化还原酶）中 B 细胞色素的功能

• 批准号: 01570142
• 负责人: KITA Kiyoshi
• 金额: $ 1.34万
• 依托单位: Juntendo University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01570142/
• 关键词: complex II; cytochrome b; mitochondria; cloning; cDNA sequence; サブユニットの解離

Complex II (succinate-ubiquinone oxidoreductase) is an important enzyme complex of both the TCA cycle and of the aerobic respiratory chain of mitochondria in eukaryotic cell and prokaryotic organisms. In this study, we have characterized the cytochrome b in complex II as described below.1) To dissociate the subunits of complex II, ionic detergent "sarkosyl" was most useful.2) In vivo reconstitution system of complex II from sdh genes separated in multiple plasmids in the Escherichia coli Cell has been established.3) cDNA library of Ascarismuscle has been constructed and positive clones for cytochorme b subunit of compleX II have been screened immunologically.4) Amino acid sequence of iron-sulfur subunit in human liver was deduced from cDNA which was isolated by immunoscreening.5) Oxidation-reduction potentials of cytochromes in muscle mitochondria of Ascaris and purified complex II were determined and compared with those in mammalian mitochondria. Oxidation-reduction potential of cytochrome b_<558> of Ascaris complex II was higher than that of bovine heart complex II.

复合物II（琥珀酸-泛醌氧化还原酶）是真核细胞和原核生物中TCA循环和线粒体有氧呼吸链的重要酶复合物。在这项研究中，我们已经如下所述表征了复合物II中的细胞色素B。1）为了解离复合物II的亚基，离子型去污剂“sarkosyl”最有效。2）建立了大肠杆菌多质粒sdh基因复合物II的体内重组体系。构建了蛔虫肌肉cDNA文库，免疫学筛选了compleX Ⅱ细胞色素B亚基的阳性克隆。5）测定了蛔虫肌线粒体和纯化的复合物II细胞色素的氧化还原电位，并与哺乳动物线粒体的氧化还原电位进行了比较。蛔虫复合物II的细胞色素B_氧化还原电位<558>高于牛心复合物II。

项目成果

Kiyoshi Kita: "Mitochondrial Encephalomyopathies" Raven Press, AC 500 (1991)

Kiyoshi Kita：“线粒体脑肌病” Raven Press，AC 500 (1991)

Kiyoshi Kita: "Human complex II(succinate-ubiquinone oxidoreductase):cDNA cloning of iron sulfur(Ip)subunit of liver mitochondria" Biochemical and biophysical research communications. 166. 101-108 (1990)

Kiyoshi Kita：“人类复合物II（琥珀酸泛醌氧化还原酶）：肝线粒体铁硫（Ip）亚基的cDNA克隆”生物化学和生物物理研究通讯。

Kiyoshi Kita: "Mitochondrial ENcephalomyopathies (in press)" Raven Press, #. 500 (1991)

Kiyoshi Kita：“线粒体脑肌病（正在印刷中）”Raven Press，

Kiyoshi Kita: Raven Press. Mitochondrial Encephalomyopathies, 500 (1991)

喜多清：乌鸦出版社。

Kiyoshi Kita: "Human comples II (succinateーubiquinone oxidoreductase); cDNA cloning of iron sulfur (Ip) subunit of liver mitochondria" Biochemical and Biophysical reserach communications. 166. 101-108 (1990)

Kiyoshi Kita：“人类复合体 II（琥珀酸泛醌氧化还原酶）；肝线粒体铁硫 (Ip) 亚基的 cDNA 克隆”《生物化学和生物物理研究通讯》166. 101-108 (1990)。