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1989 Fiscal Year Final Research Report Summary

Mechanism of Translocation of Proteins across Mitochondrial and Microsomal Membranes.

Research Project

Project/Area Number 63480503
Research Category

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

Allocation TypeSingle-year Grants
Research Field 代謝生物化学
Research InstitutionKyushu University

Principal Investigator

OMURA Tsuneo  Kyushu University, Graduate School of Medical Sciences, Department of Molecular Biology, Professor, 大学院・医学系研究科, 教授 (80029933)

Project Period (FY) 1988 – 1989
KeywordsMitochondrial membrane / Microsomal membrane / Protein translocation across membranes
Research Abstract

1. Mitochondrial targeting signals in the precursor peptides of adrenodoxin and P-450(SCC) were examined, and were found in both cases to be in the 15-20 amino acid residues-long amino terminal portions of the extension peptides.
2.20-25 amino acid residues-long amino terminal portions of the extension peptides of P-450(SCC), P-450(11beta), and adrenodoxin precursors were chemically synthesized, and used in detecting mitochondrial protein components having specific affinity to the synthetic peptides. A few mitochondrial proteins were found to have high affinity to the peptides.
3. Selective adsorption of mitochondrial protein precursors to cardiolipin-containing liposomes was found. This finding suggests some role of cardiolipin in the specific binding of protein precursors to mitochondira.
4. A metal protease which efficiently processed mitochondrial protein precursors to mature forms was purified to homogeneity from rat and bovine liver mito chondria. The purified enzyme was a heterodimer of 2 subunits, whose molecular weights were 52 KD,and 55 KD. Cleavage of adrenodoxin precursor by the purified protease confirmed its endoprotease nature.
5. Structural requirements of the amino terminal signal peptides for co-translational insertion of the peptides into microsomal membrane were examined with a cell-free system. The insertion of the peptides was depend on the hydrophobicity of the hydrophobic stretch in the signal peptides.
6. Structural requirements of stop-transfer sequences for efficient interruption of the translocation of peptides across microsomal membrane were examined.
The hydrophobicity of, the middle portion of stop-transfer sequences had primary importance in the interruption of peptide translocation. Charged amino acid residues following the hydrophobic stretch affected the efficiency of the stop-transfer sequences.

  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Wei-Jia Ou: "Specific Binding of Mitochondrial Protein Precursors to Liposomes Containing Cardiolipin." Journal of Biochemistry. 103. 589-595 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yasumitsu Takagi: "Molecular Cloning and Nucleotide Sequence of cDNA of Microsomal Carboxyesterase El of Rat Liver." Journal of Biochemsitry. 104. 801-806 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takeo Kumamoto: "Critical Region in the Extension Peptide for the Import of Cytochrome P-450(SCC) Precursor into Mitochondria." Journal of Biochemistry. 105. 72-78 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shirou Kirita: "Expression of Two Kinds of Cytochrome P-450(11β) mRNA in Bovine Adrenal Cortex." Journal of Biochemistry. 104. 683-686 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toshihide Hashimoto: "Cloning and Characterization of Bovine Cytochrome P-450(11β) Genes." Journal of Biochemistry. 105. 676-679 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Wei-Jia Ou: "Purification and Characterization of a Processing Protease from Rat Liver Mitochondria." EMBO Journal. 8. 2605-2612 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tomoyuki Harano, et al.: "Biosynthesis and Localization of Rat Liver Microsomal Carboxyesterase E1." Journal of Biochemistry, 103, 149-155 (1988).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Wei-Jia Ou, et al.: "Specific Binding of Mitochondrial Protein Precursors to Liposomes Containing Cardiolipin." Journal of Biochemistry, 103, 589-595 (1988).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yasumitsu Takagi, et al.: "Molecular Cloning and Nucleotide Sequence of cDNA of Microsomal Carboxyesterase E1 of Rat Liver." Journal of Biochemistry, 104, 801-806 (1988).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takeo Kumamoto, et al.: "Critical Region in the Extension Peptide for the Import of Cytochrome P-450 (SCC) Precursor into Mitochondria." Journal of Biochemistry, 105, 72-78 (1989).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shirou Kirita, et al.: "Expression of Two Kinds of Cytochrome P-450(11beta) mRNA in Bovine Adrenal Cortex." Journal of Biochemistry, 104, 683-686 (1988).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toshihide Hashimoto, et al.: "Cloning and Characterization of Bovine Cytochrome P-450(11beta) Genes." Journal of Biochemistry, 105, 676-679 (1989).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Osamu Hatano, et al.: "Immunohistochemical Localization of Adrenodoxin in Bovine Adrenal Cortex by Protein A-Gold Technique." Histochemistry, 91, 89-97 (1989).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Wei-Jia Ou, et al.: "Purification and Characterization of a Processing Protease from Rat Liver Mitochondria." EMBO Journal, 8, 2605-2612 (1989).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yasuki Nonaka, et al.: "Molecular Cloning and Sequence Analysis of cDNA Encoding Rat Adrenal Cytochrome P-450(11beta)." FEBS Letters, 255, 21-26 (1989).

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1993-03-26  

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