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2001 Fiscal Year Final Research Report Summary

Analysis of Structure and Function of Proteasome Derived from Rat Liver Microsome

Research Project

Project/Area Number 11480170
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Structural biochemistry
Research InstitutionHimeji Institute of Technology

Principal Investigator

KOIDE Takehiko  Himeji Institute of Technology, Faculty of Science, Department of Life Science, Professor, 理学部, 教授 (60018695)

Co-Investigator(Kenkyū-buntansha) TOKUNAGA Fuminori  Himeji Institute of Technology, Faculty of Science, Department of Life Science, Research Associate, 理学部, 助手 (00212069)
Project Period (FY) 1999 – 2001
KeywordsProteasome / ERb Proteasome / 20S Proteasome / Quality Control in the Endoplasmic reticulum / ER-Associated Degradation / Rat Liver
Research Abstract

This investigation was aimed to characterize structural and enzymic properties of the ERb proteasome which we have isolated, for the first time in the world, from rat liver microsome fraction as an ER membrane-bound proteasome and to determine the structural element specific in the ERb proteasome, and we clarified the followings :
1. On reverse-phase HPLC, in addition to β2 and α5 of 20S proteasome, ERb was found to contain extra subunits corresponding to β2 and α5 in 20S proteasome, which we referred as to B and L.
2. TWO dimension gel electrophoresis revealed that B subunit is more acidic than β2 due to phosphorylation, and L subunit was essentially the same as α5, although slight difference in mobility was observed.
3. On MALDI-TOF/MS, lysylendopeptidase digest of B showed the same pattern as that of β2.
4. N-terminal amino acid analysis revealed that α5 has Thr and that of L was blocked. Taken together with the results of MALDI-TOF/MS, The N-terminal sequence of L was suggested as Ac-Met-Phe-Leu-Thr-where Thr was the same as N-terminal of α5.
In summary, ERb contains one each of two distinctive subunits (referred to as B and L) which is absent from 20S. B is a counterpart of β2 in 20S and distinctive from β 2 in polarity and isoelectric point L is a counterpart of a5, having 3 additional residues to N-terminal of α5 and the N-terminal residue Met was acetylated. These structural feature may explain the property of ERb to bind to ER membrane.

  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] Fuminori Tokunaga: "Secretion, γ-carboxylation, and endoplasmic reficulum-associated degradation of cnimeras with mutually exechang ed Gla domain"Thrombosis Research. 99. 511-521 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sadao Wakabayashi: "Intracellular degradation of histidine-rich glycoprotein mutants: Tokushima-1 and 2 mutants are degraded by the different"Journal of Biochemistry. 128. 201-206 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Fuminori Tokunaga: "ER-associated degradation of misfolded N-linked glycoproteins is suppressed by inhibition of ER mannosidase I"Journal of Biological Chemistry. 275. 40757-40764 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 小出武比古: "論文「タンパク質も品質管理されている」書名「タンパク質分解の不思議」"株式会社クバプロ. 187 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 小出武比古: "「新生タンパク質の品質管理機構」書名「細胞の形態形成の基本メカニズム」"株式会社金芳堂. 200 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Fuminori Tokunaga: "Secretion, γ-carboxylation, and endoplasmic reticulum-associated degradation of chimeras with mutually exchanged Gla domain between human protein C and prothrombin"Thrombosis Research. 99. 511-521 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sadao Wakabayashi: "Intracellular degradation of histidine-rich glycoprotein mutants : Tokushima-1 and 2 mutants are degraded by the different proteolytic systems"Journal of Biochemistry. 128. 201-206 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Fuminori Tokunaga: "ER-associated degradation of misfolded N-inked glycoproteins is suppressed by inhibition of ER mannosidase I"Journal of Biological Chemistry. 275. 40754-40764 (2000)

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

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Published: 2003-09-17  

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