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1993 Fiscal Year Final Research Report Summary

Mechanism of Integration of Proteins into Biomembranes.

Research Project

Project/Area Number 03404071
Research Category

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

Allocation TypeSingle-year Grants
Research Field 代謝生物化学
Research InstitutionKYUSHU UNIVERSITY

Principal Investigator

OMURA Tsuneo  Kyushu University, Graduaet School of Medical Science, Professor, 大学院・医学系研究科, 教授 (80029933)

Co-Investigator(Kenkyū-buntansha) SAKAGUCHI Masao  Kyushu University, Graduaet School of Medical Science, Research Associate, 大学院・医学系研究科, 助手 (30205736)
MORIHASHI Ken-ichirou  Kyushu University, Graduaet School of Medical Science, Research Associate, 大学院・医学系研究科, 助手 (30183114)
MIHARA Katsuyoshi  Kyushu University, Graduaet School of Medical Science, Associate Professor, 大学院・医学系研究科, 助教授 (40029963)
Project Period (FY) 1991 – 1993
KeywordsMembrane proteins / Biomembranes / Membrane topologie / Membrane biogenesis
Research Abstract

The mechanism of Integration of protein molecules into endoplasmic reticulum (ER) membrane and mitochondiral (Mt) outer membrane was studied with cell-free systems and cultured cells. The import of precursor proteins into mitochondria was also studied with the cell-free system. The following results were obtained.
1. The orientation of the signal-anchor sequence at the amino-terminus in the ER membrane depends on the balance between the amino-terminal charge and the length of the following stretch of hydrophobic amino acids. It was confirmed that the amino-terminus of the signal-anchor sequence of P-450(arom) is in the luminal side of the ER membrane.
2. A highly conserved proline-rich region is present after the hydrophobic stretch of the signal-anchor sequence of microsomal P-450s. The proline-rich region was essential for the formation of the correct conformation of P-450s.
3. The structural requirements for the interruption of the transfer of growing peptides across the ER membrane were clarified. The function of the stop-transfer sequence mainly depends on the hydrophobic core sequence.
4. The peptide-translocation apparatus of the ER membrane was examined by chemically cross-linking the membrane components with the translocating nascent peptides. A novel 9 kDa membrane protein was identified as a possible component of the translocation apparatus.
5. Liver cytosol was found to stimulate the mitochondrial import of the precursor proteins synthesized by the wheat germ lysate translation system. A novel import-stimulating factor was purified from the cytosol. It was a molecular chaperon which holds the precursor proteins in the import-competent forms.

  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] Tomoko Kuroiwa: "Systematic analysis of stop-transfer sequence for microsomal membrane." J.Biol.Chem.266. 9251-9255 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masao Sakaguchi: "Yeast gene which suppresses the defect in protein export of a secY mutant of escherichia coli." J.Biochem.109. 799-802 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shigeki Furuya: "Cytosolic and mitochondrial surface factor-independent import of a synthetic peptide into mitochondria." EMBO J.10. 1759-1766 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masao Sakaguchi: "Function of signal and signal-anchor sequences are determined by the balance between the hydrophobic segment and the N-terminal charge." Proc.Natl.Acad.Sci.USA.89. 16-19 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Jun Iwahashi: "Characterization of adrenodoxin precursor expressed in Escherichia coli." J.Biochem.111. 451-455 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masao Sakaguchi: "Mitochondrial porin can be translocated across both endoplasmic reticulum and mitochondrial membranes." J.Biochem.112. 243-248 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kazuya Murakami: "An isozyme of microsomal carboxyesterase,carboxyesterase Sec,is secreted from rat liver into the blood." J.Biochem.113. 61-66 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Naomi Hachiya: "A mitochondrial import factor purified from rat liver cytosol is an ATP-dependent conformational modulator for precursor proteins." EMBO J.12. 1579-1586 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tomoko Kuroiwa: "Detection of a novel 9-kDa endoplasmic reticulum membrane protein in mammalian cells by chemical cross-linking with translocating nascent peptides." J.Biochem.114. 541-546 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Okura Shimozawa: "Core glycosylation of cytochrome P-450(arom).Evidence for localization of N-terminus of microsomal cytochrome P-450 in the lumen." J.Biol.Chem.268. 21399-21402 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Soh Yamazaki: "Importance of the proline-rich region following signal-anchor sequence in the formation of correct conformation of microsomal cytochrome P-450s." J.Biochem.114. 652-657 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tomoko Kuroiwa: "Systematic analysis of stop-trasfer sequence for microsmal membrane." J.Biol.Chem.266. 9251-9255 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masao Sakaguchi: "Yeast gene which suppresses the defect in protein export of a secY mutant of Escherichia coli." J.Biochem.109. 799-802 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shigeki Furuya: "Cytosolic and mitochondrial surface factor-independent import of a synthetic peptide into mitochondria." EMBO J.10. 1759-1766 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masao Sakaguchi: "Function of signal and signal-anchor sequences are determined by the balance between the hydrophobic segment and the N-terminal charge." Proc.Natl.Acad.Sci.USA. 89. 16-19 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Jun Iwahashi: "Characterization of adrenodoxin precursor expressed in Escherichia coli." J.Biochem.111. 451-455 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masao Sakaguchi: "Mitochondrial Porin can be translocated across both endoplasmic reticulum and mitochondrial membranes." J.Biochem.112. 243-248 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kazuya Murakami: "An isozyme of microsomal carboxyesterase, carboxyesterase Sec, is secreted from rat liver into the blood." J.Biochem.113. 61-66 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Naomi Hachiya: "A mitochondrial import factor purified from rat liver cytosol is an ATP-dependent conformational modulator for precursor proteins." EMBO J.12. 1579-1586 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tomoko Kuroiwa: "Detection of a Novel 9-kDa endoplasmic reticulum membrane protein in mammalian cells by chemical cross-linking with translocating nascent peptides." J.Biochem.114. 541-546 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Okura Shimozawa: "Core Glycosylation of cytochrome P-450(arom). Evidence for localization of N-terminus of microsomal cytochrome P-450 in the lumen." J.Biol.Chem.268. 21399-21402 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Soh Yamazaki: "Importance of the proline-rich region following signal-anchor sequence in the formation of correct conformation of microsomal cytochrome P-450s." J.Biochem.114. 652-657 (1993)

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

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Published: 1995-03-27  

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