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Posttranslational Modifications of Proteins during Intracellular Transport

Research Project

Project/Area Number 02670117
Research Category

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

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

Principal Investigator

IKEHARA Yukio  Fukuoka Univ. (Biochemistry) Professor, 医学部, 教授 (70037612)

Co-Investigator(Kenkyū-buntansha) OHKUBO Kumiko  Fukuoka Univ. (Biochemistry) Assistant, 医学部, 助手 (80223759)
TAKAMI Noboru  Fukuoka Univ. (Biochemistry) Assistant, 医学部, 助手 (80154904)
MISUMI Yoshio  Fukuoka Univ. (Biochemistry) Assistant, 医学部, 助手 (10148877)
Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1991: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1990: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsProprotein processing / Furin / Proalbumin / Complement C3 / GPI anchor / Alkaline phosphatase / Dipeptidyl peptidase IV / Enzyme defect / 細胞内プロセシング / ジペプチジルペプチダ-ゼIV / アミノ酸置換 / 活性中心 / 小胞体内蛋白質分解 / cDNAクロ-ニング / 変換酵素インヒビタ- / 補体C3前駆体
Research Abstract

1. -Identification of furin involved in the processing of proproteins
1) Primary structure of furin deduced from the cDNA : The cDNA of furin was isolated from human and rat liver cDNA libraries. The cDNA sequences indicate that the human and rat furins consist of 794 and 793 amino acids, respectively. The primary structure of each furin shows high homology to that of the yeast Kex2 protease, a prohormone-processing enzyme, and contains three potential sites for glycosylation and a transmenbrane domain at a COOH-terminal region.
2) Intracellular localization and processing activity of furin : The furin cDNA was transfected into COS-1 cells. Immunoelectron-microscopical observations revealed that furin is localized in the Golgi complex. When albumin or complement C3 was expressed by transfection of each cDNA, each proform was completely processed into a mature form by furin co-expressed in the same cells. These results indicate that furin, localized in the Golgi complex, is an endoproteas … More e involved in the processing of proforms of secretory proteins.
2. Biosynthesis, processing and transport of membrane proteins
1) GPI-anchored proteins : We have established that the ectoenzyme alkaline phosphatase (ALP) is initially synthesized as a proform having a hydrophobic sequence at the COOH terminus, which is immediately cleaved and replaced by a glycosyl-phosphatidylinositol (GPI) anchor, and the GPI-anchored form is transported to the cell surface. When the biosynthesis of GPI was completely blocked by 2-fluoro-2-deoxyglucose, the newly synthesized ALP remained in the endoplasmic reticulum as the proform, which was then gradually converted to a soluble form by proteolytic removal of the COOH-terminal segment, followed by its secretion. The results indicate that the GPI anchoring is prerequisite for ALP to be transported to the cell surface.
2) An enzyme defect due to proteolytic degradation in the endoplasmic reticulum (ER) : The ectoenzyme dipeptidyl peptidase IV (DPPIV) was found to be defective in a rat substrain. Cloning and sequencing of the DPPIV cDNA identified a point mutation which leads to substitution of Gly^<633>->Arg at the active site sequence of DPPIV. In addition, the mutant DPPIV was found to undergo rapid degradation in the ER without being transported to the cell surface. Thus, it is concluded that the rapid degradation of the mutant DPPIV in the ER results, in the enzyme defect in the affected rat. Less

Report

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

    (65 results)

All Other

All Publications (65 results)

  • [Publications] Misumi,Yoshio: "Molecular cloning and sequencing of the cDNA of rat liver α_1-protease inhibitor andits expression COS-1 cells" Journal of Biochemistry. 108. 230-234 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Misumi,Yoshio: "Nucleotide and deduced amino acid sequence of rat complement C3" Nucleic Acids Research. 18. 2178 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Oda,Kimimitsu: "Production of propeptide-directed antibody:Its application to the purification of proalbumin and analysis of proalbumin processing" Journal of Biochemistry. 108. 549-553 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Misumi,Yoshio: "Intracellular processing of complement pro-C3 and proalbumin is inhibited by rat α_1-protease inhibitor variant (Met^<352>→Arg) in transfected cells" Biochemical and Biophysical Research Communication. 171. 236-242 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Misumi,Yoshio: "Sequence of the cDNA encoding rat furin,a possible propeptide-processing endoprotease" Nucleic Acids Research. 18. 6719 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Ohkubo,Kumiko: "Molecular cloning and characterization of rat contrapsin-like protease inhibitor and related proteins" Journal of Biochemistry. 109. 243-250 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Misumi,Yoshio: "Primary structure of rat liver 5'-nucleotidase deduced from the cDNA:Presence of the COOH-terminal hydrophobic domain for possible posttranslational modification by glycophospholipid" Journal of Biological Chemistry. 265. 2178-2183 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Misumi,Yoshio: "Primary structure of human placental 5'-nucleotidase and identification of the glycolipid anchor in the mature form" Eurpean Journal of Biochemistry. 191. 563-569 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Ogata,Shigenori: "Membrane-anchoring domain of rat liver 5'-nucleotidase:Identification of the COOH-terminal serine-523 covalently attached with a glycolipid" Biochemistry. 29. 7923-7927 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Takami,Noboru: "Intracellular accumulation and oligosaccharide processing of alkaline phosphatase under disassembly of the Golgi complex caused by brefeldin A" European Journal of Biochemistry. 194. 805-810 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Oda,Kimimitsu: "Selective processing of proalbumin determined by site-specific mutagenesis" Biochemical and Biophysical Reseach Communication. 175. 690-696 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Ohkubo,Kumiko: "Cloning,structure,and expression of mouse contrapsin and its related protein" Biochemical Journal. 276. 337-342 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Oda,Kimimitsu: "Bafilomycin A1 inhibits the targeting of lysosomal acid hydroloases in cultured hepatocytes" Biochemical and Biophysical Research Communications. 178. 369-377 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Misumi,Yoshio: "Functional expression of furin demonstrating its intracellular localization and endoprotease activity for processing of proalbumin and complement pro-C3" Journal of Biological Chemistry. 266. 16954-16959 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Oda,Kimimitsu: "Sequence requirements for proteolytic cleavage of precursors with paired basic amino acids" Biochemical and Biophysical Research Communications. 179. 1181-1186 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Nagahama,Masami: "Evidence that differentiates between precursor cleavages at dibasic and Arg-X-Lys/Arg-Arg sites" Journal of Biochemistry. 110. 806-811 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Volknandt,Walter: "5'-Nucleotidase from the electric ray electri lobe:Primary structure and relation to mammalian and procaryotic enzymes" European Journal of Biochemistry. 202. 855-861 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Casanova,James E.: "Direct apical sorting of rat liver dipeptidylpeptidase IV expressed in Madin-Darby canine kidney cells" Journal of Biological Chemistry. 266. 24428-24432 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Takami,Noboru: "Aberrant processing of alkaline phosphatase precursor caused by blocking the synthesis of glycosyl-phosphatidylinositol" Journal of Biological Chemistry. 267. 1042-1047 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Ogata,Shigenori: "Identification of the active site residues in dipeptidyl peptidase IV by affinity labeling and site-directed mutagenesis" Biochemistry. 31. (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tsuji,Emiko: "Missense mutation (Gly^<633>→Arg) of dipeptidyl peptidase IV causes its retention and rapid degradation in the endoplasmic reticulum" Journal of Biological Chemistry.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Misumi, Y., et al.: "Molecular cloning and sequencing of the cDNA of rat liver alpha_1-protease inhibitor and its expression in COS-1 cells" J. Biochem. 108. 230-234 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Misumi, Y., et al.: "Nucleotide and deduced amino acid sequence of rat complement C3" Nucleic Acids Res.18. 2178-2178 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Oda, K., et al.: "Production of propeptide-directed antibody : Its application to the purification of proalbumin and analysis of proalbumin processing" J. Biochem.108. 549-553 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Misumi, Y., et al.: "Intracellular processing of complement pro-C3 and proalbumin is inhibited by rat alpha_1-protease inhibitor variant (Met^<352>-> Arg) in transfected cells" Biochem. Biophys. Res. Commun.171. 236-242 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Misumi, Y., et al.: "Sequence of the cDNA encoding rat furin, a possible propeptideprocessing endoprotease" Nucleic Acids Res.18. 6719-6719 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Misumi, Y., et al.: "Primary structure of rat liver 5'-nucleotidase deduced from the cDNA : Presence of the COOH-terminal hydrophobic domain for possible posttranslational modification by glycosylphosphatidylinositol" J. Biol. Chem.265. 2178-2183 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Misumi, Y., et al.: "Primary structure of human placental 5'-nucleotidase and identification of the glycolipid anchor in the mature form" Eur. J. Biochem.191. 563-569 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Ogata, S., et al.: "Membrane-anchoring domain of rat liver 5'-nucleotidase : Identification of the COOH-terminal serine-523 covalently attached with a glycolipid" Biochemistry. 29. 7923-7927 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Takami, N., et al.: "Intracellular accumulation and oligosaccharide processing of alkaline phosphatase under disassembly of the Golgi complex caused by brefeldin A" Eur. J. Biochem.194. 805-810 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Ohkubo, K., et al.: "Molecular cloning and characterization of rat contrapsinlike protease inhibitor and related proteins" J. Biochem. 109. 243-250 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Oda, K., et al.: "Selective processing of proalbumin determined by sitespecific mutagenesis" Biochem. Biophys. Res. Commun.175. 690-696 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Ohkubo, K., et al.: "Cloning, structure, and expression of mouse contrapsin and a related protein" Biochem. J.276. 337-342 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Oda, K., et al.: "Bafilomycin Al inhibits the targeting of lysosomal acid hydrolases in cultured hepatocytes" Biochem. Biophys. Res. Commun.178. 369-377 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Misumi, Y., et al.: "Functional expression of furin demonstrating its intracellular localization and endoprotease activity for processing of proalbumin and complement pro-C3" J. Biol. Chem.266. 16954-16959 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Oda, K., et al.: "Sequence requirements for proteolytic cleavage of precursors with paired basic amino acids" Biochem. Biophys. Res. Commun.179. 1181-1186 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Nagahama, M., et al.: "Evidence that differentiates between precursor cleavages at dibasic and Arg-X-Lys/Arg-Arg sites" J. Biochem.110. 806-811 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Volknandt, W., et al.: "5'-Nucleotidase from the electric ray electric lobe : Primary structure and relation to mammalian and procaryotic enzymes" Eur. J. Biochem.202. 855-861 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Casanova, J. E., et al.: "Direct apical sorting of rat liver dipeptidyl peptidase IV expressed in Madin-Darby canine kidney cells" J. Biol. Chem.266. 24428-24432 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Takami, N., et al.: "Aberrant processing of alkaline phosphatase precursor caused by blocking the synthesis of glycosyl-phosphatidylinositol" J. Biol. Chem.267. 1042-1047 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Ogata, S., et al.: "Identification of the active site residues in dipeptidyl peptidase IV by affinity labeling and site-directed mutagenesis" Biochemistry. 31, March issue.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tsuji, E., et al.: "Missense mutation (Gly^<633>->Arg) of dipeptidyl peptidase IV causes its retention and rapid degradation in the endoplasmic reticulum" J. Biol. Chem.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Ohkubo,Kumiko: "Molecular cloning and characterization of rat contrapsin-like protease inhibitor and related proteins" journal of biochemistry. 109. 243-250 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Oda,Kimimitsu: "Selective processing of proalbumin determined by site-specific mutagenesis" Biochemical and Biophysical Research Communications. 175. 690-696 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Ohkubo,Kumiko: "Cloning,structure and expression of cDNA for mouse contrapsin and a related protein" Biochemical Journal. 276. 337-342 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Oda,Kimimitsu: "Bafilomycin Al inhibits the targeting of lysosomal acid hydrolases in cultured hepatochtes" Biochemical and Biophysical Research Communications. 178. 369-377 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Misumi,Yoshio: "Functional expression of furin demonstrating its intracellular localization and endoprotease activity for processing of proalbumin and complement pro-C3" Journal of Biological Chemistry. 266. 16954-16959 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Oda,Kimimitsu: "Sequence requirements for proteolytic cleavage of precursors with paired basec amino aicds" Biochemical and Biophysical Research Communications. 179. 1181-1186 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Nagahama,Masami: "Evidence that differentiates between precursor cleavages at debasic and Arg-X-Lys/Arg-Arg sites" Journal of Biochemistry. 110. 806-811 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Volknandt,Walter: "5'-Nucleotidase from the electric ray electric lobe:Primary structure and relation to mammalian and procaryotic enzymes" European Journal of Biochemistry. 202. 855-861 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Casonova,James E.: "Direct apical sorting of rat liver dipeptidylpeptidase IV expressed in Madin-Darby canine Kidney cells" Journal of Biological Chemistry. 266. 24428-24432 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Takami,Noboru: "Aberrant processing of alkaline phosphatase precursor caused by blocking the synthesis of glycosyl-phosphatidylinositol" Journal of Biological Chemistry. 267. 1042-1047 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Ogata,Shigenori: "Identification of the active site residues in dipeptidyl peptidase IV by affinity babeling and site-directed mutagenesis" Biochemistry. 31. (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Misumi,Yoshio: "Molecular cloning and sequencing of the cDNA of rat liver α_1ーprotease inhibitor and its expression in COSー1 cells" Journal of Biochemistry. 108. 230-234 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Misumi,Yoshio: "Nucleotide and deduced amino acid sequence of rat complement C3" Nucleic Acids Research. 18. 2178 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Oda,Kimimitsu: "Production of propeptideーdirected antibody:Its application to the purification of proalbumin and analysis of proalbumin processing" Journal of Biochemistry. 108. 549-553 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Misumi,Yoshio: "Intracellular processing of complement proーC3 and proalbumin is inhibited by rat α_1ーprotease inhibitor variant(Met^<352>→Arg)in transfected cells" Biochemical and Biophysical Research Communication. 171. 236-242 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Misumi,Yoshio: "Sequence of the cDNA encoding rat furin,a possible propeptideーprocessing endoprotease" Nucleic Acids Research. 18. 6719 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Ohkubo,Kumiko: "Molecular cloning and characterization of rat contrapsinーlike protease inhibitor and related proteins" Journal of Biochemistry. 109. 243-250 (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Misumi,Yoshio: "Primary structure of rat liver 5'ーnucleotidase deduced from the cDNA:Presence of the COOHーterminal hydrophobic domain for possible posttranslational modification by glycophospholipid" Journal of Biological Chemistry. 265. 2178-2183 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Misumi,Yoshio: "Primary structure of human placental 5'ーnucleotidase and identification of the glycolipid anchor in the mature form" European Journal of Biochemistry. 191. 563-569 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Ogata,Shigenori: "Membraneーanchoring domain of rat liver 5'ーnucleotidase:Identification of the COOHーterminal serineー523 covalently attached with a glycolipid" Biochemistry. 29. 7923-7927 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Takami,Noboru: "Intracellular accumulation and oligosaccharide processing of alkaline phosphatase under disassembly of the Golgi complex caused by brefeldin A" European Journal of Biochemistry. 194. 805-810 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Oda,Kimimitsu: "Selective processing of proalbumin determined by siteーspecific mutagenesis" Biochemical and Biophysical Research Communication. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Ohkubo,Kumiko: "Cloning,structure,and expression of mouse contrapsin and its related protein" Biochemical Journal. (1991)

    • Related Report
      1990 Annual Research Report

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

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