| Project/Area Number |
05044176
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
OMURA Tsuneo Kyushu University, Graduate School of Medical Science, Professor Emeritus, 大学院医学系研究科, 名誉教授 (80029933)
大村 恒男 (1994) 九州大学, 大学院・医学系研究科, 名誉教授
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| Co-Investigator(Kenkyū-buntansha) |
CHERNOGOLOV アレクロイ エー. ベラルーシ科学アカデミー, 生物有機化学研究所, 研究員
USANOV Sergey a. Academy of Sciences of Belarus, Institute of Bioorganic Chemistry, Professor, 生物有機化学研究所, 教授
NEGISHI Masahiko National Institute of Environmental Health Sciences, Investigator, 研究員
MIHARA Katsuyoshi Kyushu University, Graduate School of Medical Science, Associate Professor, 大学院医学系研究科, 助教授 (40029963)
HARA Takayuki Nakamura Gakuen University, Faculty of Home Economics, Associate Professor, 家政学部, 助教授 (10164998)
CHERNOGOLOV Alexey a. Academy of Sciences of Belarus, Institute of Bioorganic Chemistry, Research Asso
ALEXEY A. Ch ベラルーシ科学アカデミー, 生物有機化学研究所, 研究員
SELGEY A. Us ベラルーシ科学アカデミー, 生物有機化学研究所, 教授
相良 康弘 高知医科大学, 医学部, 助教授 (60162319)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1994: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1993: ¥4,000,000 (Direct Cost: ¥4,000,000)
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| Keywords | Steroids / CytochromeP450 / P450scc / Adrenodoxin / 構造活性相関 / P-450SCC |
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| Research Abstract |
Several forms of microsomal and mitochondrial cytochrome P450s catalyze the metabolism of steroids in animal tissues. The object of this research project is to elucidate the regulatory mechanism of steroid metabolism by the P450s on the basis of the structure-function relationship of the P450s and their reducing systems. Intimate collaboration between the Japanese investigators and the two foreign research groups, Dr.Negishi's group in USA and Dr.Usanov's group in Belarus, was planned and carried out. Dr.Usanov and Dr.Chernogolovvisited Fukuoka to do research with the Japanese investigators. Dr.Hara visited Minsk and Research Triangle Park to do research with Dr.Usanov and Dr.Negishi, respectively.
Major effort of the research was focused on the mechanism of reduction of steroid-metabolizing mitochondrial P450s by NADPH via NADPH-adrenodoxin reductase and adrenodoxin. P450scc and adrenodoxin were the major subjects of the research. The topology of P450scc in the inner mitochondrial membrane was studied by the use of site-specific antibodies, and the major portion of P450scc molecule was shown to be exposed on the matrix-side surface of the membrane. The interaction between P450scc and adrenodoxin was studied by cross-linking them with various chemical cross linkers and by the use of radio-labeled adrenodoxin. In order to study the interaction, site-directed mutagenesis of adrenodoxin was carried out. The native and the mutated adrenodoxins were expressed in E.coli, and the expressed proteins were purified and used in the kinetical studies on the electron transfer from adrenodoxin to P450scc. It was also found that the precursor of adrenodoxin was efficiently expressed in E.coli, and that the expressed precursor contained the iron-sulfur active center and could reduce P450scc in a reconstituted in vitro system.
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