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Regulation of gene expression and function of cytochrome b5

Research Project

Project/Area Number 09044092
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research Field Functional biochemistry
Research InstitutionKochi University

Principal Investigator

YUBISUI Toshitsugu  Kochi University, Department of Biology, Professor, 理学部, 教授 (00019564)

Co-Investigator(Kenkyū-buntansha) ポーター トッドD.  ケンタッキー大学, 薬学部, 助教授
ステッグルス アランW.  ノースイースタンオハイオ大学, 医学部, 教授
キャンベル ウイルバーH  ミシガン工科大学, 生命科学部, 教授
PORTER Todd D.  University of Kentucky, Department of Pharmacy, Associate professor
CAMPBELL Wilbur H.  Michigan Technological University, Life Science, Professor
STEGGLES Alen W.  Northeastern Ohio Medical Universtiy, Department of Molecular Biology, Professor
キャンベル ウイルパーH  ミシガン工科大学, 生命科学部, 教授
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1998: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1997: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsCytochrome b5 / cDNA cloning / gene expression / tunicate / co-expression / recombinant protein / ホルモン代謝 / シトクロムb_5 / 遺伝子 / 発現調節 / 機能解析 / タンパク質発現
Research Abstract

In these studies we investigated the regulation of gene expression and function of cytochrome b5.
1. Yubisui has succeeded in cloning 2 cDNAs from tunicates. One of them obtained from Polyandrocarpa misakiensis has about 1.8 kbp including the coding regions of 405 bp for 135 amino acids, and the other from Ciona savignyi has about 790 bps including the coding region of 402 bp for 134 amino acids. Both amino acid sequences have about 75% homology to those of mammalian cytochronie b5s.
2. Steggles also cloned a cDNA for swine cytochrome bS, which contained a 24 bp insertion after 96th codon to code for G1u97, Pro98 and stop99 to produce the soluble form of cytochrome b5. Regulation of gene expression of the soluble and membrane-bound form depends on the functin of promotors.
3. Porter constructed a co-expression system for cytochrome b5, P450, and P450 reductase in Eschericlzia coli. Cytochrome b5 effectively accerelated the drug metabolism by cytochrome P450/P450 reductase system.
4. Campbell investigated the function of cytochrome b5 moiety in nitrate reductase. The cytochrome b5 and flavoprotein domains of the nitrate reductase were expressed as a fusion protein, and crystalyzed. Based on the crystalyzed structure he succeeded in changing the NADPH-dependent enzyme to NADH-specific enzyme by site-directed mutagenesis.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] Yanase, T.et al.: "Immunohistochemical Study on Cytochrome b5 in Human Adrenal Gland and in Adrenocortical Adenomas from Patient with" Endocrine Journal. 45(1). 89-95 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shiraishi, N.et al.: "Engeneering of pyridine nucleotide specificity of nitrate reductase: Mutagenesis of recombinant cytochrome b reductase fragment of Neu" Arch・Biochem.Biophys.358. 104-115 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] VanDenMrak, P.K. et al.: "The isolation and characterization of the soluble and membrane-bound porfcine cytochrome b5 cDNAs." Biochem. Biophys. Res. Commun.240. 80-83 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ratnam, K.et al.: "Spectroscopic and kinetic characterization of the recombinant cytochrome c reductase of nitrate reductse : identification of the rate" J.Biol.chem.272. 2122-2128 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Tang, Y.: "Deletion Analysis of the FAD Domain of NADPH-Cytochrome P450 Reductase" Flavins and Flavoproteins. 1996. 479-482 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shiraishi, N., Campbell, W.H.: "Expression of Nitrate Reductase FAD-Containing Fragments in Pichia" Flavins and Flavoproteins. 1996. 931-934 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yanase, T.et al.: "Immunohistochemical Stusy on Cytochrome b5 in Human Adrenal Gland and in Adrenocortical Adenomas from Patient with Cushing's Syndrome" Endocrine Journal. 45(1). 89-95 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shiraishi, N.et al., Croy, C., Kaur, J.and Campbell, W.H.: "Engeneering of pyridine nucleotide specificity of nitrate reductase : Mutagenesis of recombinant cytochrome b reductase fragment of Neurospora crassa NADPH : nitrate reductase." Arch.Biochem.Biophys.358. 104-115 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yoshida, S.et al.: "Analysis by Fourier Transform Infrared Spectroscopies of Protein Structures of Soluble NADH-Cytochrome b5 Reductases Prepared by Site-directed Mutagenesis." Biospetroscopy. 3. 215-223 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] VanDerMrak, P.K.et al.: "The isolation and characterization of the soluble and membrane-bound porcine cytochrome b5 cDNAs." Biochem.Biophys.Res.Commun.240. 80-83 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Tang, Y.: "Deletion Analysis of the FAD Domain of NADPH-Cytochrome P450 Reductase." In Flavins and Flavoproteins(eds.Massey, V.and Williams, Jr., C.H.)1996,479-482, Univ.of Calgary Press, Calgary, Canada. (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ratnam, K., Shiraishi, N.Campbell, W.H.and Hille, R.: "Spectroscopic and kinetic characterization of the recombinant cytochrome c reductase of nitrate reductse : identification of the rate limiting catalytic step." J.Biol.chem.272. 2122-2128 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shiraishi, N., Campbell, W.H.: "Expression of Nitrate Reductase FAD-Containing Fragments in Pichia" In Flavins and Flavoproteins(eds.Massey, V.and Williams, Jr., C.H.)pp.931-934, Univ.of Calgary Press, Calgary, Canada. (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Su W., J.A.Mertens, K.Kanamaru, Wilbur H.Campbell, and N.M.Crawford.: "Analysis of wild-type and mutant plant nitrate reductase expressed in the methylotrophic yeast Pichia pastoris." Plant Physiology. 115. 1135-1143 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yanase,T.et al.: "Immunohistochemical Study on Cytochrome b5 in Human Adrenal Gland and in Adrenocortical Adenomas from atient with" Endocrine Journal. 45(1). 89-95 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shiraishi,N.et al.: "Engeneering of pyridine nucleotide specificity of nitrate reductase:Mutagenesis of recombinant cvtochrome v reductase fragment of New" Arch.Biochem.Biophys.358. 104-115 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] VanDenMrak,PK.et al.: "The isolanon and characterization of the soluble and membranebound porcine cvtochrome b5 cDNAs." Biochem.Biophys.Rse.Commun.240. 80-83 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Ramam,K.et al.: "Spectroscopic and kinetic characterization of the recombinant cvtochrome c reductase of nitrate reductse:identification of the rate lin." J.Biol.chem.272. 2122-2128 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Tang,Y.: "Deletion Analysis of the FAD Domain of NADPH-Cytochrome P450 Reductase." Flavins and Flavoproteins. 1996. 479-482 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shiraishi,N.,Campbell,W.H.: "Expression of Nitrate Reductase FAD-Containing Fragments in Pichia" Flavins and Flavoproteins. 1996. 931-934 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yanase, T.et al.: "Immunohistochemical Study on Cytochrome b5 in Human Adrenal Gland and in Adrenocortical Adenomas from" Endocrine Journal. 45(1). 89-95 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Yoshida, S.et al.: "Analysis by Fourier Transform Infrared Spectroscopies of Protein Structures of Soluble NADH-Cytochrome b5 Reductases" Biospectroscopy. 3. 215-223 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Tang, Y.: "Deletion Analysis of the FAD Domain of NADPH-Cytochrome P450 Reductase" Flavins and Flavoproteins. 479-482 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] VanDerMrak, P.K.et al.: "The isolation and characterization of the soluble and membrane-bound porcine cytochrome b5 cDNAs" Biochem.Biophys.Res.Commum.240. 80-83 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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