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Transport of Proteins into Mitochondria

Research Project

Project/Area Number 02454147
Research Category

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

Allocation TypeSingle-year Grants
Research Field General medical chemistry
Research InstitutionKumamoto University

Principal Investigator

MORI Masataka  Kumamoto University Medical School, Professor, 医学部, 教授 (40009650)

Co-Investigator(Kenkyū-buntansha) MORI Shunsuke  Kumamoto University Medical School, Assistant Professor, 医学部, 助手 (70230068)
MORI Kaoru (MURAKAMI)  Kumamoto University Medical School, Assistant Professor, 医学部, 助手 (20230063)
滝口 正樹  熊本大学, 医学部, 講師 (40179578)
Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥6,900,000 (Direct Cost: ¥6,900,000)
Fiscal Year 1991: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1990: ¥4,200,000 (Direct Cost: ¥4,200,000)
KeywordsPrecursor proteins / Intracellular transport / Mitochondrial biogenesis / Cytosolic factors / PBF (presequence binding factor) / Ornithine transcarbamylase / Hsp70 / Presequence / Presequence binding factor(PBF)
Research Abstract

Ornithine transcarbamylase (OTC), a mitochondrial matrix enzyme of the urea cycle, is initially synthesized on cytosolic free ribosomes as a larger precursor pOTC with an NH_2-terminal presequence of 32 amino acid residues. pOTC is released into a cytosolic pool and is imported into the mitochondrial matrix with a half life of 1-2 min. We established an in vitro import system in which the purified pOTC was imported into isolated mitochondria. Using this system, we showed that a cytosolic protein factor (s) in the reticulocyte lysate is required for the import of the purified pOTC. A protein factor that binds to pOTC but not to mature OTC and was named presequence binding factor (PBF), was purified from the lysate. The purified PBF migrated as a single polypeptide of about 50, 000 Da on SDS-PAGE. On sucrose gradients, pOTC and PBF cosedimented as a complex of 7S. The purified PBF markedly stimulated the import of the purified pOPC into the mitochondria. PBF-stimulated pOTC import was further enhanced by hsp70 purified from yeast ; the hsp70 alone had little effect. Thus, PBF binds to the presequence portion of the precursor and may hold it in a transport-competent form in cooperation with hsp70. We next asked whether PBF is also involved in transport of other mitochondrial proteins. The precursor for aspartate aminotransferase synthesized in the PBF-depleted lysate failed to be imported into the mitochondria. Readdition of the purified PBF to the depleted lysate fully restored the import. Therefore, PBF appears to be involved in transport of a set of mitochondrial proteins having presequences.

Report

(3 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • Research Products

    (21 results)

All Other

All Publications (21 results)

  • [Publications] Arakawa,Hiroyuki: "NH_2-terminal 14-16 amino acids of mitochondrial and bacterial thiolases can direct mature ornithine carbamoyltransrerase into mitochondria" J.Biochem.107. 160-164 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Amaya,Yoshihiro: "Isolation of a yeast gene,SRHI,that encodes a homologue of the 54K subunit of mammalian signal recognition particle" J.Biochem.107. 457-463 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Murakami,Kaoru: "Presequence does not prevent folding of a purified miochondrial precursor protein and is essential for association with a reticulocyte cytosolic factor(s)" J.Biochem.108. 207-214 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Murakami,Kaoru: "Purified presequence binding factor (PBF) forms an import-competent complex with a purified mitochondrial precursor protein" EMBO J.9. 3201-3208 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Haraguchi,Yougo: "Cloning and sequence of a cDNA incoding human carbamyl phosphate synthetase" Gene. 107. 335-340 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Murakami,Kaoru: "Presequence binding factor (PBF)-dependent and -independent import of proteins into mitochondria"

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Mori,Masataka(分担執筆): "Fatty Acid Oxidation:Clinical,Biochemical,and Molecular Aspects" Alan R.Liss, 7 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Arakawa, H. . Amaya, Y., and Mori, M.: "NH_2-terminal 14-16 amino acids of mitochondrial and bacterial thiolases can direct mature ornithine carbamoyltransferase into mitochondria" J. Biochem.107. 160-164 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Amaya, Y., Nakano, A., Ito, K., and Mori, M.: "Isolation of a yeast gene, SRHI, that encodes a homologue of the 54K subunit of mammalian signal recognition particle" J. Biochem.107. 457-463 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Murakami, K., Tokunaga, F., Iwanaga, S., and Mori, M.: "Presequence does not prevent folding of a purified mitochondrial precursor protein and is essential for association with a reticulocyte cytosolic factor (s)" J. Biochem.108. 207-214 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Murakami, K., and Mori, M.: "Purified presequence binding factor (PBF) forms an import-competent complex with a purified mitochondrial precursor protein" EMBO J.9. 3201-3208 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Haraguchi, Y. Uchino, T., Takiguchi, M., Endo, F., Mori, M., and Matsuda, I.: "Cloning and sequence of a cDNA encoding human carbamyl phosphate synthetase I : molecular analysis of hyperammonemia" Gene. 107. 335-340 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Murakami, K., Tanase, S., Morino, Y., and Mori, M.: "Presequence binding factor (PBF)-dependent and -independent import of proteins into mitochondria"

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Haraguchi,Yougo: "Cloning and sequence of a cDNA incoding human carbamyl phosphate synthetase" Gene. 107. 335-340 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Murakami,Kaoru: "Presequence binging factor(PBF)ーdependent andーindependent import of proteins into mitochondria"

    • Related Report
      1991 Annual Research Report
  • [Publications] 森 正敬: "タンパク質のミトコンドリア移行と異常" BIOmedica. 7. 139-143 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Arakawa,Hiroyuki: "NH_2ーterminal 14ー16 amino acids of mitochondrial and bacterial thiolases can direct mature ornithine carbamoyltransferase into mitochondria" J.Biochem.107. 160-164 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Amaya,Yoshihiro: "Isolation of a yeast gene,SRHI,that encodes a homologue of the 54K subunit of mammalian signal recognition particle" J.Biochem.107. 457-463 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Murakami,Kaoru: "Presequence does not prevent folding of a purified mitochondrial precursor protein and is essential for association with a reticulocyte cytosolic factor(s)" J.Biochem.108. 207-214 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Murakami,Kaoru: "Purified presequence binding factor (PBF) forms an importーcompetent complex with a purified mitochondrial precursor protein" EMBO J.9. 3201-3208 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Mori,Masataka(分担執筆): "Fatty Acid Oxidation: Clinical,Biochemical,and Molccular Aspects" Alan R.Liss, 7 (1990)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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