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Molecular mechanism of functional expression of the motor protein involved in endoplasmic streaming in Characean cells.

Research Project

Project/Area Number 07458167
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionNAGOYA UNIVERSITY

Principal Investigator

FUJIME Sugie Higashi  Nagoya University, School of Science, Associate professor, 大学院・理学研究科, 助教授 (60022662)

Co-Investigator(Kenkyū-buntansha) KOHAMA Kazuhiro  Gunma University, Medical School, Professor, 医学部, 教授 (30101116)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1996: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1995: ¥5,300,000 (Direct Cost: ¥5,300,000)
KeywordsChara / endoplasmic streaming / motor protein / sliding movement / myosin family / cloning / アクチン系運動 / 滑り運動機構
Research Abstract

The motor protein responsible for cytoplasmic streaming in Characean cells (Chara myosin) can slide actin filaments at the velocity of 60mum/s, which is ten times faster than the sliding movement of skeletal muscle myosin. We succesfully purified this Chara motor protein, Chara myosin, heavily contaminated proteases, and found some distinct characterstics of this myosin, in addition to the fastest sliding movement, and reported the results (reference 1).
In order to get clue for elucidating the molecular basis of this distinct function of Chara myosin, we examined by using various techniques, particularly molecular biology techniques for cloning and sequencing of Chara myosin.
It was very difficult to purify mRNA from Characean cells, which contained endoplasm only 1% of the total cell volume. However, we improved the mRNA purification method and fortunately got mRNA with sufficient purity, although quantity was very small. We made cDNA library for cloning of the fastest Chara myosin. Th … More e probes for screening of the cDNA library were obtained from PCR using primers designed according to peptide microsequences of the Chara myosin and also consensus sequences of various myosins. The product of PCR contained 5 clones, each of which showed great similarity to any of known myosins. We are now continuing to analyze the clones obtained and the structure of Chara myosin.
So far, the sliding velocity is understood to correlate with the actomyosin ATPase activity. We found, however, that the velocity did not correlated with the actomyosin ATPase activity, but strongly correlated with the reciprocal of the ATPase activity of myosin itself, which corresponds to the life time of the intermediate state of the rate limitting step of the myosin ATPase cycle, myosin ADP Pi. The longer the life time of myosin ADP Pi, the faster the sliding velocity. Therefore, we can predict that the fastest Chara myosin must have much longer life time at the intermediate state than any other myosins. This finding can promisingly lead us to the clue to elucidate the molecular mechanism of the tremendously fast velocity of Chara myosin. Less

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Sugie Higashi-Fujime: "The fastest actin-based motor protein from the green algae, Chara, and its distinct mode of interaction with actin." FEBS Letters,. 375. 151-154 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sugie Higashi-Fujime,: "The mechanism for mechanochemical energy transduction in actin-myosin interaction revealed by in vitro motility assay with ATP analogues." Biochem. Biophys. Res. Commun.221. 773-778 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tetsu Hozumi: "Maleimidobenzoyl actin : Its biochemcal properties and in vitro motility." J. Biochem. (Tokyo). 119. 151-156 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Akio Nakamura: "Calcium inhibition oa actin-myosin interaction." Proc. Yamada Conference. 39. 270-276 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sugie Higashi-Fujime.Ryoki Ishikawa, Hidenori Iwasawa, Osamu Kagami, Eiji Kurimoto, Kazuhiro Kohama, and Tetru Hozumi: "'The fastest actin-based motor protein from the green algae, Chara, and its distinct mode of interaction with actin.'" FEBS Letters. 375. 151-154 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sugie Higashi-Fujime, and Tetsu Hozumi: "'The mechanism for mechanochemical energy transduction in actin-myosin interaction revealed by in vitro motility assay with ATP analogues.'" Biochem.Biophy.Res.Commun.221. 773-778 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tetsu Hozumi, Masao Miki and Sugie Higashi-Fujime: "'Maleimidobenzoyl actin : Its biochemical properties and in vitro motility'" J.Biochem.119. 151-156 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Akio Nakamura and Kazuhiro Kohama: "'Calcium inhibition oa actin-myosin interaction.'" Proc.Yamada Conference. 39. 270-276 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sugie Higashi-Fujime: "The fastest actin-based motor protein from the green algae,Chara,and its distinct mode of interaction with actin." FFBS Letters. 375. 151-154 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Sugie-Higashi-Fujime: "The mechanism for mechanochemical energy transduction in actin-myosin interaction revealed by in vitro motility assay with ATP analogues." Biochem.Biophys.Res.Commun.221. 773-778 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Tetsu Hozumi: "Maleimidobenzoyl actin:Its biochemcal properties and in vitro motility." J.Biochem.(Totyo). 119. 151-156 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Akio Nakamura: "'Calcium inhibition oa actin-myosin interaction.'" Proc.Yamada Conference. 39. 270-276 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] S.Higashi-Fujime: "The fastest actin-based motor protein from the green algae,Chara,and its distinct mode of itneraction with actin." FEBS Lett.375. 151-154 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] T.Hozumi: "Maleimidobenzoyl actin : its biochemical properties and in vitro motility." J.Biochem.119. 151-156 (1996)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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