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Yeast Flavohemoglobin from Candida norvegensis: Its Structural, Spectral, and Stability Properties

Research Project

Project/Area Number 12640659
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 動物生理・代謝
Research InstitutionFUKUSHIMA MEDICAL UNIVERSITY (2001)
Tohoku University (2000)

Principal Investigator

MATSUOKA Ariki  Fukushima Med. Univ., Schl. Of Med., Associate Professor, 医学部, 助教授 (30222293)

Co-Investigator(Kenkyū-buntansha) TAJIMA Gen-ichi  Tohoku Univ., Grad. Schl. Of Sci., Research Assistant, 大学院・理学研究所, 助手 (00197360)
SHIKAMA Keiji  PHP Laboratory, Director, 所長 (40004337)
OCHLAI Takehiko  Fukushima Med. Univ., Schl. Of Med., Professor, 医学部, 教授 (40075031)
Project Period (FY) 2000 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2001: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2000: ¥2,700,000 (Direct Cost: ¥2,700,000)
Keywordshemoglobin / myoglobin / yeast / autoxidation / high-level expression system / two domain structure / flavin / acid-catalysis
Research Abstract

A hemoglobin-like protein from yeast, Candida norvegensis, is composed of a single polypeptide chain with 389 amino acid residues, but of two distinct domains carrying different functions: One is a heme-containing oxygen binding domain in the N-terminal region and the other is a FAD containing reductase domain found in the remaining C-terminal region (Iwaasa, H., Takagi, T. and Shikama, K. (1992) J. Mol. Biol. 227, 948-954; Shikama, K., Matsuoka, A. and Iwaasa, H. (1995) Int. J. Biochem. CellBiol. 27, 1107-1115).
To investigate the structure and function of this yeast flavohemoglobin in more detail, we have constructed two expression systems using a vector PMAL-c2 in E. coli cells (TB-1 strain). One is for the whole protein of Candida flavohemoglobin, and the other for the oxygen binding domain (heme domain) alone. The resulting recombinant proteins were then purified using gel filtration on Sephadex G-75 followed by DEAE-cellulose chromatography, and were examined for their spectral and stability properties in veiw of the oxygen-binding function.

Report

(3 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] Korenaga, S.: "A primitive myoglobin from Tetrahymena pyriformis : Its heme environment, autoxidizability, and genomic DNA structure"Biochim.Biophys.Acta. 1543. 131-145 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Suzuki, T.: "Dual nature of the distal histidine residue in the autoxidation reaction of myoglobin and hemoglobin : comparison of the H64 mutants"Eur.J.Biochem.. 267. 6166-6174 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 四釜慶治: "ミオグロビン・ヘモグロビンの自動酸化反応-その分子機構と結合酸素の安定性-"生物物理. 234. 74-79 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 落合威彦: "呼吸調節の系統発生:特にconstant relative alkalinityについて"LiSA. 8. 686-692 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Yasuda, J P.: "The α1β1 contact of human hemoglobin plays a key role in stabilizing the bound dioxygen : Further evidence from the iron valency hybrids"Eur.J.Biochem.. 269. 202-211 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shikama, K.: "Human hemoglobin and its heme oxidation : A new paradigm for oxygen stabilization involving the α1β1 contact"Trends in Comparative Biochemistry and Physiology. (In press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Matsuoka, A.: "Proceedings of the XIIth International Conference on Invertebrate O_2 Binding Proteins"Yeast Flavohemoglobin from Candida norvegensis : Its Structural, Spectral, and Stability Properties(In press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Korenaga, S.: "A primitive myoglobin from Tetrahymena pyriformis: Its heme environment autoxidizability and genomic DNA structure"Biochim Biophys. Acta. 1543. 131-145 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Suzuki, T: "Dual nature of the distal histidine residue in the autozidation reaction of myoglobin and hemoglobin: comparison of the H64 mutants"Eur. J. Biochem. 267. 6166-6174 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shikama, K.: "The Molecular Mechanism of Autoxidation for Myoglobin and Hemoglobin Seibutsu Butsuri"234. 74-79 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ochiai, T.: "Phyiogeny in the control of respiration with special reference to the constant relative alkalinity"LiSA. 8. 686-692 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Yasiida, J P.: "The α1β1 contact of human hemoglobin plays a key role in stabilizing the bound dioxygen: Further evidence from the iron valency hybrids"Eur. J. Biochem. 269. 202-211 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shikama,K.: "Human hemoglobin and its heme oxidation: A new paradigm for Oxygen stabilization involving the α1β1 contact"Trends in Comparative Biochemistry and Physiology. (in press). (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Matsuoka, A.: "Yeast Flavohemoglobin from Candida norvegensis: Its Strultural, Spectral, and Stability Properties"Proceedings of the XIIth International Conference on Invertebrate O_2 Binding Proteins (in press). (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Korenaga, S.: "A primitive myoglobin from Tetrahymena pyriformis : Its heme environment, autoxidizability, and genomic DNA structure"Biochim. Biophys. Acta. 1543. 131-145 (2000)

    • Related Report
      2001 Annual Research Report
  • [Publications] Suzuki, T.: "Dual nature of the distal histidine residue in the autoxidation reaction of myoglobin and hemoglobin : comparison of the H64 mutants"Eur. J. Biochem.. 267. 6166-6174 (2000)

    • Related Report
      2001 Annual Research Report
  • [Publications] 四釜慶治: "ミオグロビン・ヘモグロビンの自動酸化反応 -その分子機構と結合酸素の安定性-"生物物理. 234. 74-79 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 落合威彦: "呼吸調節の系統発生:特にconstant relative alkalinityについて"LiSA. 8. 686-692 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Yasuda, J P.: "The α1β1 contact of human hemoglobin plays a key role in stabilizing the bound dioxygen : Further evidence from the iron valency hybrids"Fur. J. Biochem.. 269. 202-211 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Shikama, K.: "Human hemoglobin and its heme oxidation : A new paradigm for oxygen stabilization involving the α1β1 contact"Trends in Comparative Biochemistry and Physiology. (In press). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Matsuoka, A.: "Yeast Flavohemoglobin from Candida norvegensis : Its Structural, Spectral, and Stability Properties"Proceedings of the XIIth International Conference on Invertebrate O_2 Binding Proteins. (In press). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Korenaga,S.: "A primitive myoglobin from Tetrahymena pyriformis : Its heme environment, autoxidizability, and genomic DNA structure."Biochim.Biophys.Acta. 1543. 131-145 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Suzuki,T.: "Dual Nature of the Distal Histidine Residue in the Autoxidation Reaction of Myoglobin and Hemoglobin : Comparison of the H64 Mutants."Eur.J.Biochem.. 267. 6166-6174 (2000)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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