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1998 Fiscal Year Final Research Report Summary

Mechanism for insertion of tail-anchored proteins into the endoplasmic reticulum

Research Project

Project/Area Number 09680704
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Cell biology
Research InstitutionKansai Medical University

Principal Investigator

MASAKI Ryuichi  Kansai Medical University, Faculty of Medicine, Assistant Professor, 医学部, 講師 (70140283)

Co-Investigator(Kenkyū-buntansha) YAMAMOTO Akitsugu  Faculty of Medicine, Assistant Professor, 医学部, 講師 (30174775)
Project Period (FY) 1997 – 1998
KeywordsHPC-1 / syntaxin 1A / Tail-anchored protein / ER integration / Intracellular transport
Research Abstract

HPC-1/syntaxin lA (HPC-l), which plays an important role in vesicular transport to the plasma membrane, possesses a hydrophobic sequence at its C terminus, In the present study, we investigated the role of the C-terminal portion of HPC-1 in intracellular localization. When expressed from cDNA in COS cells, wild-type HPC-1 was localized in the Golgi complex and the plasma membrane. HPC-l localized in the Golgi complex was chased away to the plasma membrane in the presence of cycloheximide. Truncation of the hydrophobic domain resulted in the cytoplasmic localization of the mutant, thus indicating that the domain indeed functions as a membrane anchor. The fusion protein with the N-terminal region of bovine opsin (the 0P3 extension), which contains the N-glycosylation sites, was glycosylated in transfected COS cells. In addition, we found that a chimeric protein consisting of Escherichia coil maltose binding protein with the last 24 amino acids of HPC-1 and the 0P3 extension is also glycosylated and localized along the exocytic pathway of transfected cells similar to HPC- 1. Taken together, these indicate that HPC-1 has a transmembrane structure, and suggest that the protein is first inserted into the endoplasmic reticulum and then transported to the plasma membrane via the exocytic pathway. In addition, our results indicate that the C-terminal portion of HPC-1is important for its intracellular localization.

  • Research Products

    (18 results)

All Other

All Publications (18 results)

  • [Publications] Kadota, Y.: "A newly identified membrance proteins localized exclusively in intracellular organilles of neurons." Mol. Brain Res.46. 265-273 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Komada, M.: "Hrs, a tyrosin kinase substrate with a con served double zinc finger domain, is licalized to the cyteplasmic surface" J. Biol. chem.272. 20538-20544 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamamoto, A.: "Bafilomycin Al prevents mutation of airtip hagic vacuoles by inhibiting fusion between autophagos ones and lysosomes" Cell Struct. Funct.23. 33-42 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masaki, R.: "Important roles of the c-terminal portion of HPC-1/syntaxin/A in menbrane anchorine and intracellular" J. Biochem.124. 311-318 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamamoto, A.: "Studies on intracellular structures of COS cells by X-ray microscopy." J. Synchrot. Rad.5. 1105-1107 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tagawa, Y.: "A 60 KDa Plasma membrane protein changes its localization to autophagosomes and autolysosome membranes." Cell Struct. Funct.印刷中. (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yoshida, H.: "Comfensated endocytosis of LDL by hamster cells co-expressing the two distinct mutant LDL receptors defectire in endecytosis and ligan finds" J. Lipid Res.印刷中. (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masaki、R.: "Membrane Proteins : Structure, function and expression control" 九州大学出版, Karger AG, 19 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 柾木龍一: "タンパク質化学" 広川書店 印刷中, (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kadota, Y.: "A newly identified membrane protein localized exclusively in intracellular organelles of neurons." Molecular Brain Research. 46. 265-273 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Komada, M.: "Hrs, a tyrosine kinase substrate with a conserved double zinc finger domain, is localized to the cytoplasmic surface of early endosomes." J.Biol.Chem. 272. 20538-20544 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamamoto, A.: "Bafilomycin A1 prevents mutation of autophagic vacuoles by inhibiting fusion between autophagosomes and lysosomes in rat hepatoma cell line, H-4-II-E cells." Cell Struct.Funct. 23. 33-42 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masaki, R.: "Important roles of the C-terminal portion of HPC-1/syntaxin 1A in membrane anchoring and intracellular localization." J.Biochem. 124. 311-318 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamamoto, A.: "Studies on intracellular structures of COS cells by X-ray microscopy." J.Synchrot.Rad.5. 1105-1107 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tagawa, Y.: "A 60 kDa plasma membrane protein changes its localization to autophagosome and sutolysosome membranes during induction of autophagy in rat hepatoma cell line, H-4-II-E cells." Cell Struct.Funct.(in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoshida, H.: "Compensated endocytosis of LDL by hamster cells co-expressing the two distinct mutant LDL receptors defective in endocytosis and ligand binding." J.Lipid, Rrs.(in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masaki, R.: Microsomal aldehyde dehydrogenase in the endoplasmic reticulum, in "Membrane proteins : Structure, function and expression control".ed.by Mihara, K and Hamasaki, N.Kyushu Umiversity Press, Fukuoka/S.Karger AG,Basel, 19 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masaki, R.: Aldehyde Dehydrogenase, in "Protein Chemisrty". (in Japanese). Hirokawa Shuppan (in press), (1999)

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

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Published: 1999-12-08  

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