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Gene Analysis of Photosystem II complex for Its 3-Dimensional Structure Elucidation

Research Project

Project/Area Number 12640641
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 植物生理
Research InstitutionRIKEN-The Institute of Physical and Chemical Reseach

Principal Investigator

SHEN Jianren  RIKEN, The Institute of Physical and Chemical Research, Photosynthesis Research Laboratory, Senior Research Scientist, 光合成科学研究室, 先任研究員 (60261161)

Project Period (FY) 2000 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2001: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2000: ¥1,800,000 (Direct Cost: ¥1,800,000)
KeywordsPHOTOSYSTEMII / PHOTOSYNTHESIS / CRYSTALSTRUCTURE / MEMBRANEPROTEINS / CRYSTALLIZATION / OXYGENEVOLUTION / SYNECHOCOCCUS VULCANUS / 立体構造
Research Abstract

The purpose of this research is to analyze genes coding for all of the subunits of oxygen-evolving photosystem (PSII) complex purified from a thermophilic cyanobacterium Thermosynechococcus vulcaus (formly Synechococcus vulcanus). We have succeeded in crystallizing the PSII complex of T vulcanus. In order to analyze the structure of PSII at atomic resolution, it is essential to know the complete composition of the complex utilized for crystallization, and the sequences of all of the subunits contained in the complex ; both of which have not been determined previously. By combining eiectrophoresis, measurement of oxygen-evolving activity, TOF-MASS measurement, and N-terminal sequencing results reported previously, we determined that there are at least 13 trans-membrane subunits and 3 extrinsic proteins existing in the PSII complex utilized for crystallization and also after crystallization. Genes for some of these components have not been cloned from T vulcanus. However, since the whole genomc sequences of another closely related thermophilic cyanobacterium T elongatLts were determined by Kazusa DNA Research Institute and will be published shortly, we focused our research on the analysis of crystal structure of PSII in the latter part of this project. By screening many heavy atom derivates, we found several derivates that yielded effective phase information ; with these information, we analysed the crystal structure of PSII at 3.7 A resolution. The resulted structure contained new information that was not available in the 3.8 A of PSII reported by Witt et al. in 2001 for PSII of T elongates. For example, the extrinsic 12 kDa protein was newly assigned in our structure, and one of the ligands to the Mn-cluster was suggested to be the C-terminal of the Dl subunits. In addition, more detailed information was obtained concerning the extrinsic loops of CP47 and CP43 that possibly protrudes into the lumenal side, the arrangements of electron transfer cofactors, etc.

Report

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

    (21 results)

All Other

All Publications (21 results)

  • [Publications] Shen J.-R., Kamiya N.: "Crystallization and the crystal properties of the oxygen-evolving photosystem II from Synechococcus vulcanus"Biochemistry. 39・48. 14739-14744 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Enami I., Yoshihara S., Tohri A., Okumura A., Ohta H., Shen J.-R.: "Cross-reconstitution of various extrinsic proteins and photosystem II complexes from cyanobacteria, red algae and higher plants"Plant and Cell Physiology. 41・12. 1354-1364 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Al-khaldi S., Coker J., Shen J.-R., Burnap R.L.: "Characterization of site-directed mutations in the manganese-stabilizing protein of Synechocystis sp. PCC 6803 unable to grow photoautotrophically in the absence of cytochrome c-550"Plant Mol. Biol.. 43・1. 33-41 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Katoh H., Itoh S., Shen J.-R., Ikeuchi M.: "Functional analysis of psbV and a novel c-type cytochrome gene psbV2 of the thermophilic cyanobacterium Thermosynechococcus elongatus strain BP-1"Plant and Cell Physiology. 42・6. 599-607 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tohri A., Suzuki T., Ohta H., Shen J.-R., Yamamoto Y., Enami I., et al.: "Comparison of the structure of the extrinsic 33 kDa protein from different organisms"Plant and Cell 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] Kamiya N., Shen J.-R.: "Crystal structure analysis of photosystem II from Thermosynechococcus vulcanus"PS2001 Proceedings, 12^<th> International Congress on Photosynthesis. CISRO Publishing (Online and CD). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Nakata Y., Satoh A., Okumura A., Ohta H., Shen J.-R., Enami I., et al.: "Distribution of extrinsic proteins among various organisms as an index of evolution of oxygen-evolving PSII"PS2001 Proceedings, 12^<th> International Congress on Photosynthesis. CISRO Publishing (Online and CD). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Suzuki T., Akiyama A., Tomo T., Ohta H., Shen J.-R., Enami I., et al.: "Reconstitution of the extrinsic 23 and 17 kDa proteins with spinach PSII which had been exchanged for the 33 kDa protein from different plant species"PS2001 Proceedings, 12^<th> International Congress on Photosynthesis. CISRO Publishing (Online and CD). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Enami I., Iwai M., Akiyama A., Nakada Y., Ohta H., Shen J.-R.: "Cross-reconstitution of cytochrome c-550 and 12 kDa protein between cyanobacterial and red algal PSII"PS2001 Proceedings, 12^<th> International Congress on Photosynthesis. CISRO Publishing (Online and CD). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] ShenJ.-R. and Kamiya N.: "Crystallization and the crystal properties of the oxygen-evolving photosystem II from Synechococcus vulcanus"Biochemistry. 39-48. 14739-14744 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Enami I., Yoshihara S., Tohri A., Okumura A., Ohta H. and Shen J.-R: "Cross-reconstitution of various extrinsic proteins and photosystem II complexes from cyanobacteria, red algae and higher plants"Plant Cell Physiol.,. 41-12. 1354-1364 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Al-Khaldi S., CokerJ., Shen J.-R. and R.L. Burnap: "Characterization of site-directed mutations in the manganese- stabilizing protein of Synechocystis sp.PCC 6803 unable to grow photoautotrophically in the absence fo cytochrome c-550"Plant Mol. Biol.. 43-1. 33-41 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Katoh H., Itoh S., Shen J.-R. and Ikeuchi M: "Functional analysis of psbV and a novel c-type cytochrome gene psbV2 of the thermophilic cyanobacterium Thermosynechococcus elongates strain BP-1"Plant Cell. Physiol.. 42-6. 599-607 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tohri A., Suzuki T., Okuyama S., Kamino K., Motoki A., Hirano M., Ohta H., Shen J.-R., Yamamoto Y. and Enami I.: "Comparison of the structure of the extrinsic 33 kDa protein from different organisms"Plant Cell Physiol.,. (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] Kamiya N. and Shen J.-R.: "Crystal structure analysis of photosytem II from Thermosynechococcus vulcanus"PS2001 Proceedings, 12'" International Congress on Photosynthesis (Online and CD), CISRO PUBLISHING, Australia. (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Nakata Y., Satoh A., Nakamura K., Okumura A., Ohta H., Shen J.-R. and Enami I.: "Distribution of extrinsic proteins among various organisms as an index of evolution of oxygen-evolving PSII"PS2001 Proceedings, 12'" International Congress on Photosynthesis (Online and CD), CISRO PUBLISHING, Australia. (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Suzuki T., Akiyama A., Iwai M., Tohri A., Tomo T., Ohta H., Shen J.-R. and Enami I.: "Reconstitution of the extrinsic 23 and 17 kDa proteins with spinach PSII which had been exchanged for the 33 kDa protein from different plant species"PS2001 Proceedings, 12^<th> International Congress on Photosynthesis (Online and CD), CISRO PUBLISHING, Australia. (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Enami L, Iwai M., Akiyama A., Nakada Y., Ohta H. and Shen J.-R.: "Cross-reconstitution of cytochrome c-550 and 12 kDa protein between cyanobacterial and red algal PSII"PS2001 Proceedings, 12^<th> International Congress on Photosynthesis (Online and CD), CISRO PUBLISHING, Australia. (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Katoh, H., Itoh, S., Shen, J.-R., Ikeuchi, M.: "Functional Analysis of psbV and a Novel c-type Cytochrome Gene psbV2 of the Thermophilic Cyanobacterium Thermosynechococcus elongatus Strain BP-1"Plat and Cell Physiology. 42・6. 599-607 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tohri, A., Suzuki, T., Okuyama, S., Kamino, K., Motoki, A., Hirano, M., Ohta, O., Shen, J.-R., Yamamoto, Y., Enami, I.: "Comparison of the structure of the extrinsic 33 kDa protein from different organisms"Plat and Cell Physiology. (In press). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Shen J.-R.,Kamiya N.: "Crystallization and the crystal properties of the oxygen evolving photosystem II from Synechococcus vulcanus."Biochemistry. 39・48. 14739-14744 (2000)

    • Related Report
      2000 Annual Research Report

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

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