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Structure-Activity Relation of p-450 and Related Enzymes

Research Project

Project/Area Number 05044176
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research InstitutionKYUSHU UNIVERSITY

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

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)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
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)
KeywordsSteroids / CytochromeP450 / P450scc / Adrenodoxin / 構造活性相関 / P-450SCC
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.

Report

(2 results)
  • 1994 Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (44 results)

All Other

All Publications (44 results)

  • [Publications] Shin-ichirou Honda: "Ad4BP regulating steroidogenic P-450 gene is a member of steroid hormone receptor superfamily." J.Biol.Chem.268. 7494-7502 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ken-ichirou Morohashi: "Activation of CYP11B gene promoters by the steroidogenic cell-specific transcription factor,Ad4BP." Mol.Endocrinol.7. 1196-1204 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Masatoshi Nomura: "Three forms of rat CYP11B genes:11B-hydroxylase gene,aldosterone synthase gene,and a novel gene." J.Biochem.113. 144-152 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Soh Yamazaki: "Importance of the proline-rich region following signal-anchor sequence in the formation of correct conformation of microsomal cytochrome P-450s." J.Biochem.114. 652-657 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Okura Shimozawa: "Core glycosylation of cytochrome P-450(arom).Evidence for localization of N terminus of microsomal cytochrome P-450s in the Lumen." J.Biol.Chem.268. 21399-21402 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kazuya Murakami: "The transmembrane region of microsomal cytochrome P450 identified as the endoplasmic reticulum retention signal." J.Biochem.116. 164-175 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ken-ichirou Morohashi: "Functional difference between Ad4BP and ELP,and their distributions in steroidogenic tissues." Mol.Endocrinol.18. 643-653 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Osamu Hatano: "Sex-dependent expression of a transcription factor,Ad4BP,regulating steroidogenic P-450 genes in the gonads during prenatal and postnatal rat development." Development. 120. 2787-2797 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Takanori Yasukochi: "Putative functions of phenylalaine-350 of pseudomonas putida cytochrome P-450cam." Biochim.Biophys.Acta. 1204. 84-90 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kazuhide Nakamura: "Significant contribution of arginine-112 and its positive charge of pseudomonas putida cytochrome P-450CAM in the electron transport from putidaredoxin." Biochim.Biophys.Acta. 1207. 40-48 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yoshiyuki Sakai: "In-vitro evidence for the regulation of 17,20-lyase activity by cytochrome b_5 in adrenocortical adenomas from parients with cushing's syndrome." Clin.Endocrinol.40. 205-209 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yoshiyuki Sakai: "Mechanism of abnormal production of adrenal androgens in parients with adrenocortical adenomas and carcinomas." J.clin.Endocrinol.Metab.78. 36-40 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Iwasaki: "Inherent versatility of P450 oxygenase:conferring dehydroepiandrosterone hydroxylase activity to P450 2a-4 by a single amino acid mutation at position 117." J.Biol.Chem.269. 9079-9083 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] D.Schwarz: "Rotation of cytochrome P450scc(CYPllAl)in proteoliposomes studied by delayed fluorescence depolarization." Biochem.Biophys.Res.Cummuns.195. 889-896 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] A.A.Chernogolov: "Selective chemical modification of Cys^<264> with diiodofluorescein iodacetamide as a tool to study the membrane topology of cytochrome P450scc(CYPllAl)." FEBS Letters. 340. 83-88 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] A.A.Chernogolov: "Investigation of the membrane topolology of cytochrome P450scc." Futura. 9. 53-55 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Masao Sakaguchi: "Frontiers in Biotransformation,Vol.8" Akademie Verlag, 253 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Tsuneo Omura: "Handbook of Experimental Pharmacology,Vol.105:Cytochrome P450" Springer-Verlag Berlin, 350 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shin-ichirou Honda: "Ad4BP regulating steroidogenic P-450 gene is a member of steroid hormone receptor superfamily." J.Biol.Chem.268. 7494-7502 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ken-ichirou Morohashi: "Activation of CYP11B gene promoters by the steroidogenic cell-specific transcription factor, Ad4BP." Mol.Endocrinol.7. 1196-1204 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Masatoshi Nomura: "Three forms of rat CYP11B genes : 11B-hydroxylase gene, aldosterone synthase gene, and a novel gene." J.Biochem.113. 144-152 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Soh Yamazaki: "Importance of the proline-rich region following signal-anchor sequence in the formaion of correct conformation of microsomal cytochrome P-450s." J.Biochem.114. 652-657 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Okura Shimozawa: "Core glycosylation of cytochrome P-450 (arom). Evidence for localization of N terminus of microsomal cytochrome P-450s in the lumen." J.Biol.Chem.268. 21399-21402 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kazuya Murakami: "The transmembrane region of microsomal cytochrome P450 identified as the endoplasmic reticulum retention signal." J.Biochem.116. 164-175 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ken-ichirou Morohashi: "Functional difference between Ad4BP and ELP,and their distributions in steroidogenic tissues." Mol.Endocrinol.18. 643-653 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Osamu Hatano: "Sex-dependent expression of a transcription factor, Ad4BP,regulating steroidogenic P-450 genes in the gonads during prenatal and postnatal rat development." Development. 120. 2787-2797 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Takanori Yasukochi: "Putative functions of phenylalaine-350 if pseudomonas putida cytochrome P-450cam." Biochim.Biophys.Acta. 1204. 84-90 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kazuhide Nakamura: "Significant contribution of arginine-112 and its positive charge of pseudomonas putida cytochrome P-450CAM in the electron transport from putidaredoxin." Biochim.Biophys.Acta. 1207. 40-48 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yoshiyuki Sakai: "In-vitro evidence for the regulation of 17,20-lyase activity by cytochrome b_5 in adrenocortical adenomas from parients with cushing's syndrome." Clin.Endocrinol.40. 205-209 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yoshiyuki Sakai: "Mechanism of abnormal production of adrenal androgens in parients with adrenocortical adenomas and carcinomas." J.Clin.Endocrinol.Metab.78. 36-40 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Iwasaki: "Inherent versatility of P450 oxygenase : conferring dehydroepiandrosterone hydroxylase activity to P450 2a-4 by a single amino acid mutation at position 117." J.Biol.Chem.269. 9079-9083 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] D.Schwarz: "Rotation of cytochrome P450scc (CYP11A1) in proteoliposomes studied by delayd fluorescence depolarization." Biochem.Biophys.Res.Cummuns. 195. 889-896 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] A.A.Chernogolov: "Selective chemical modification of Cys^<264> with diiodofluorescein iodacetamide as a tool to study the membrane topology of cytochrome P450scc(CYP11A1)." FEBS Letters. 340. 83-88 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] A.A.Chernogolov: "Invesitigation of the membrane topology of cytochrome P450scc." Futura. 9. 53-55 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Masao Sakaguchi: Akademie Verlag. Frontiers in Biotransformation, Vol.8, 253 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Tsuneo Omura: Springer-Verlag Berlin. Handbook of Experimental Pharmacology, Vol.105 : Cytochrome P450, 350 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shin-ichrou Honda: "Ad4BP regulating steroidogenic P-450 gene is a member of steroid hormone receptor superfamily." J.Biol.Chem.268. 7494-7502 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Ken-ichirou Morohashi: "Activation of CYP11B gene promoters by the steroidogenic cell-specific transcription factor,Ad4BP." Mol.Endocrinol.7. 1196-1204 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Masatoshi Nomura: "Three forms of rat CYP11B genes:11B-hydroxylase gene,aldosterone synthase gene,and a novel gene." J.Biochem.113. 144-152 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Soh Yamazaki: "Importance of the proline-richregion following signal-anchor sequence in the formation of correct conformation of microsomal cytochrome P-450s." J.Biochem.114. 652-657 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Okura Shimozawa: "Core glycosylation of cytochrome P-450(arom).Evidence for localization of N terminus of microsomal cytochrome P-450 in the lumen." J.Biol.Chem.268. 21399-21402 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] D.Schwartz: "Rotation of cytochrome P-450SCC(CYP11Al)in proteoliposomes studied by delayed fluoregeence depollarization." Biochem.Biophys.Res.Communs.195. 889-896 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Masao Sakaguchi: "Frontiers in Biotransformation,Vol.8" Topology and biogenesis of microsomal cytochrome P-450s., 253 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Tsuneo Omura: "Handbook of Experimental Pharmacology,Vol.105:Cytochrome P450" Localization of cytochrome P450 in membranes:mitochondria., 350 (1993)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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