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Physiological studies of evolutionaly conserved-perchloric acid soluble-protein (PSP) from rat liver

Research Project

Project/Area Number 10680614
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Functional biochemistry
Research InstitutionKagoshima University (1999)
The University of Tokushima (1998)

Principal Investigator

OKA Tatsuzo (1999)  Kagoshima University, Faculty of Agriculture, Professor, 農学部, 教授 (50116795)

名取 靖郎 (1998)  徳島大学, 医学部, 教授 (30035381)

Co-Investigator(Kenkyū-buntansha) 岡 達三  鹿児島大学, 農学部, 教授 (50116795)
Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1999: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1998: ¥2,100,000 (Direct Cost: ¥2,100,000)
KeywordsRat / Perchloric acid-soluble protein / Ribocuclease / mRNA / Translation / Single strand / 過塩素酸可溶性蛋白質 / PSP / 大腸菌 / 遺伝子組換え / 蛋白質合成阻害 / GSR融合蛋白質
Research Abstract

Rat liver perchloric acid-soluble protein (L-PSP) is a potent inhibitor of cell-free protein synthesis; however, its mechanism of action is not unknown. Here we show that the protein is a unique ribonuclease and that this activity is responsible for the inhibition of translation. The addition of perchloric acid-soluble protein to a rabbit reticulocyte cell-free system at a concentration of 6.2 μM led to an almost complete inhibition of protein synthesis. The kinetics are unlike those of hemin-controlled inhibitor, a protein that acts at the initiation step. The inhibition appears to be an endoribonucleolytic activity of perchloric acid-soluble protein because L-PSP directly affects mRNA template activity and induces disaggregation of the reticulocyte polysomes into 80S ribosomes, even in the presence of cycloheximide. These effects were observed with authentic as well as recombinant L-PSP. Analysis by thin-layer chromatography of [α -ィイD132ィエD1P]UTP-labeled mRNA incubated with the protein showed production of the ribonucleotide 3'-monophosphates Ap, Gp, UP and Gp, providing direct evidence that the protein is an endoribonuclease. When either 5'- or 3'-32P-labeled 5S rRNA was the substrate, L-PSP cleaved phosphodiester bonds only in the single strand region of the molecule.

Report

(3 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • Research Products

    (21 results)

All Other

All Publications (21 results)

  • [Publications] 岡 達三: "タンパク質合成阻害タンパク質PSP"生化学. 70巻. 217-221 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] asagi, a., oka, T. et al.: "Purification, characterization and differentation-dependent expression of a perchlonic acid soluble protein from rat kidney"Nephron. 79. 80-90 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Oka, T., Nishimoto, Y. et al.: "Production of functional rat liver PSP protein in Escherichia coli"Cell Mol. Life Sci.. 55. 131-134 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Carugo, KD., oka, T. et al.: "Crystallilzation and preliminary X-ray diffraction studies of perchlonic acid soluble protein (PSP) from rat liver"Acta Crystallographica. D55. 667-668 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sasagawa, T., Oka, T. et al.: "Analysis of the fatty acid components in a perchloric acid-soluble protein"Biochim. Biophys. Acta. 1437. 317-324 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Morishita, R., kawagoshi, A., et al.: "Ribonuclease activity of rat perchloric acid-soluble protein, a potent inhibitor of protein synthesis"J. Biol. Chem.. 274. 20688-20692 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T.Oka: "Translation inhibitor protein, PSP."SEIKAGAKU. 70. 217-221 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K.Asagi, T.Oka, K.Arao, I.Suzuki, MK.Thakur, K.Izumi and Y.Natori: "Purification, characterization and differntiation-dependent expression of a perchloric acid soluble protein from rat kidney"Nephron. 79. 80-90 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T.Oka, Y.Nishimoto, T.Sasagawa, H.Kanouchi, Y.Kawasaki and Y.Natori: "Production of functional rat liver PSP protein in Escherichia coli"Cell. Mol. Life Sci.. 55. 131-134 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] KD.Carugo, M.Saraste and T.Oka: "Crystallization and Preliminary X-ray diffraction studies of perchloric acid soluble protein (PSP) from rat liver"Acat Crystallographica. D55. 667-668 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T.Sasagawa, T.Oka, A.Tokumura, Y.Nishimoto, S.Munoz, M.Kuwahata, M.Okita, H.Tsuji and Y.Natori: "Analysis of the fatty acid components in a perchloric acid-soluble protein"Biochim.Biophys.Acta. 1437. 317-324 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] R.Morishita, A.Kawagoshi, T.Sawasaki, K.Madin, T.Ogasawara, T.Oka and Y.Endo: "Ribonuclease activity of rat liver perchloric acid-soluble protein, a potent inhibitor of protein synthesis"J. Biol. Chem.. 274. 20668-20692 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 岡 達三: "タンパク質合成阻害タンパク質PSP"生化学. 70巻. 217-221 (1998)

    • Related Report
      1999 Annual Research Report
  • [Publications] Asagi,A.,Oka,T.et al.: "Purification,characterization and differntiation-dependent expression of a perchioric acid soluble protein from rat kidney."Nephron. 79. 80-90 (1998)

    • Related Report
      1999 Annual Research Report
  • [Publications] Oka,T.,Nishimoto,Y.et al.: "Production of functional rat liver PSP protein in Escherichia coli."Cell Mol.Life Sci.. 55. 131-134 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Carugo,KD.,Oka,T.et al.: "Crystallizatiojn and preliminary X-ray difffaction studies of perchloric and soluble protein (PSP) from rat liver."Acta Crystallographica. D55. 667-668 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Sasagawa,T.,,Oka,T.et al.: "Analysis of the fatty acid components in a perchloric acid-soluble protein"Biochim.Biophys.Acta. 1437. 317-324 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Morishita,R.,Kawagoshi,A.et al.: "Ribonuclease activity of rat perchloric acid-soluble protein,a potent inhibitor of protein synthesis."J.Biol.Chem.. 274. 20688-20692 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Oka,T.,Nishimoto,Y.et: "Production of functional rat PSP protein in Esherichia coli." Cell.Mol.Life.Sci.(CMLS). 55(1). 131-134 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Carugo,KD.,Oka,T.et al.: "Crystallization and preliminary X-ray diffraction studies of perchloric acid soluble protein(PSP)from rat liver." Acta Crystallographica. (印刷中).

    • Related Report
      1998 Annual Research Report
  • [Publications] Sasagawa,T.,Oka,T.et al.: "Analysis of the fatty acid components in a perchloric acid-soluble protein." Biochim.Biophys.Acta. (印刷中).

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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