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Biochemical and molecular biological studies of electron partitioning among enzymes involved in the assimilation of inorganic compounds in plants

Research Project

Project/Area Number 05454019
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 植物生理
Research InstitutionOsaka University

Principal Investigator

HASE Toshiharu  Osaka University, Institute for Protein Research, Professor, たんぱく質研究所, 教授 (00127276)

Co-Investigator(Kenkyū-buntansha) FUJITA Yuichi  Osaka University, Institute for Protein Research, Research Associate, たんぱく質研究所, 助手 (80222264)
IDEGUCHI Takashi  Osaka University, Institute for Protein Research, Research Associate, たんぱく質研究所, 助手 (60203121)
Project Period (FY) 1993 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1995: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1993: ¥5,300,000 (Direct Cost: ¥5,300,000)
KeywordsFerredoxin / Glutamate synthase / Sulfite reductase / Photosynthesis / Electron transfer / 光合成電子伝達 / フェレドキシン-NADP^+還元酵素
Research Abstract

1) Using heterologous expression system in E.coli, maize ferredoxin (Fd) was site-directly mutagenized. The substituted residues were designed surrounding [2Fe-2S] cluster and on the surface of molecule. The redox potential and electron transfer abilities to Fd-dependent enzymes, such as Fd-NADP reductase and sulfite reductase, were investigated and the result showed that Fd had a specific region to interact with the each enzyme.
2) Two types of Fd isoproteins, photosynthetic and non-photosynthetic one, were expressed in a cyanobacterium, Plectonema boryanum, whose Fd gene (pet F) had been deleted. The transformant with photosynthetic Fd grew in photoautotrophic condition under light as the wild-type cells, while the transformant was not able to grow heterotrophically under dark. The transformant with non-photosynthetic Fd could not be selected probably due to the inability of this type of Fd to complement the Pet F.
3) Genes for two glutamate synthase (GOGAT), Fd-GOGAT and NADH-GOGAT,were cloned from P.boryanum. The two GOGAT genes were separately inactivated by insertion of kanamycin cartridge. The resulting mutants were found to lack the corresponding enzymatic activity. However, the remarkable change f phenotype was not observed except for an increased sensitivity to photoinhibition. The two GOGATs are probably functionally compatible.

Report

(4 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • 1993 Annual Research Report
  • Research Products

    (18 results)

All Other

All Publications (18 results)

  • [Publications] 長谷俊治: "酸化還元代謝反応から見たプラスチドの多様性" 植物細胞工学. 5. 378-386 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 長谷俊治: "電子キャリアー蛋白質の機能特性の改変" 生産と技術. 47. 47-49 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 長谷俊治: "鉄・硫黄電子伝達蛋白質から見た植物細胞の無機物還元同化系" 蛋白質核酸酵素. 40. 282-292 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 長谷俊治: "フェレドキシンとフェレドキシン依存性酵素間の分子間認識と電子伝達" 日本応用酵素協会誌. 30. 11-18 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T. Matsumura: "Non-photosynthetic type ferredoxin from maize retards photoautotophic growth of the cyanobacterium Plectonema boryanum." photosynthesis : from Light to Biosphere(ed. P. Mathis). vol. 2. 709-712 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T. Ideguchi: "cDNA cloning and functional expression of ferredoxin-dependent sulfite erductase from maize in E. coli cells." photosynthesis : from Light to Biosphere(ed. P. Mathis). vol. 2. 713-716 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T.Hase: "Diversity of oxidoreductive metabolic pathways in plastids." Plant Cell Technology. 5. 378-386 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T.Hase: "Modification of functional characteristics of an electron carrier protein." Production and Technology. 47(1). 47-49 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T.Hase: "Iron-sulfur electron transfer protein and assimilation of inorganic compounds in plants." Protein, Nucleic Acid, Enzyme. 40. 282-292 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T.Matsumura, Y.Fujita and T.Hase: "Non-photosynthetic type ferredoxin from maize retards photoautotrophic growth of the cyanobacterium Plectonema boryanum." Photosynthesis : from Light to Biosphere (ed.P.Mathis), Kluwer Academic Publishers. vol.2. 709-712

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T.Ideguchi, T.Akashi, Y.Onda and T.Hase: "cDNA cloning and functional expression of ferredoxin-dependent sulfite reductase from maize in E.coli cells." Photosynthesis : from Light to Biosphere(ed.P.Mathis)Kluwer Academic Publishers. vol.2. 713-716

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T.Matsumura: "Non-photosynthetic type ferredoxin from maize retards photoautotrophic growth of the cyanobacterium Plectonema boryanum" photosynthesis : from Light to Biosphere (ed.P.Mathis). 2. 709-712 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] T.Ideguchi: "cDNA cloning and expression of ferredoxin-dependentsulfite reductase from maize in E.cole celles" Photosynthesis : from Light to Biosphere (ed.P.Mathis). 2. 713-716 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Y.Fujita: "Identification of chl B gene and the gene product essential for the light-independent biosynthesis of chlorophyll in the cyanobacterium P.boryanam" Plant Cell Physial.in press (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] 長谷俊治: "フェレドキシンとフェレドキシン依存性酵素間の分子間認識と電子伝達" 日本応用酵素協会誌. 30. 11-18 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 長谷俊治: "鉄・硫黄電子伝達蛋白質から見た植物細胞の無機物還元同化系" 蛋白質核酸酵素. 40. 282-292 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 長谷俊治: "電子キャリアー蛋白質の機能物性の改変" 生産と技術. 47. 47-49 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 長谷俊治: "酸化還元代謝反応からみたプラスチドの多様性" 植物細胞工学. 5. 378-386 (1993)

    • Related Report
      1993 Annual Research Report

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

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