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Structure and function of proteasome in higher plants

Research Project

Project/Area Number 05454014
Research Category

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

Allocation TypeSingle-year Grants
Research Field 植物生理
Research InstitutionChiba University

Principal Investigator

NAKAGAWA Hiroki  Chiba University Faculty of Horticulture, Professor, 園芸学部, 教授 (70009330)

Co-Investigator(Kenkyū-buntansha) SATO Takahide  Chiba University Faculty of Horticulture Asociate Professor, 園芸学部, 助教授 (60125929)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥7,300,000 (Direct Cost: ¥7,300,000)
Fiscal Year 1994: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1993: ¥6,400,000 (Direct Cost: ¥6,400,000)
Keywords26S Proteasome / Protease / spinach leaves
Research Abstract

The 26S proteasome complex catalyzing ATP-dependent breakdown of ubiquitin-ligated proteins was purified from spinach leaves to near homogeneity by chromatography on DEAE-cellurose, gel filtration on Biogel A-1.5m and glycerol density gradient cenrifugation. The purified enzyme was shown to degrade multi-ubiquitinated, but not unmodified, lysozyme in an ATP-dependent fashion, coupled with ATPase activity supplying energy for proteolysis, and isopeptidase activity to generate free ubiquitin. By non-denaturing electrophoresis, the puried enzyme was separated into to distinct forms of the 26S complex, named 26Salpha and 26Sbeta proteasomes, with different electorophoretic mobilities. The 26S proteasome was found to consist of multiple polypeptides with molecular masses of 23-35 and 39-115kDa, which were thought to be those of a 20S proteasome with multicatalytic proteinase activity and an associated regulatory part with ATPase and de-ubiquitinating activities, respectively. the subunit multiplicity of the spinach 26S proteasome dosely resembled that of rat liver with minor differences in certain components. No sulfhydryl bond was involved in the assembly of this multi-component polypeptide complex. Electron microscopy showed that the 26S proteasome complex had a "caterpillar" like shape, consisting of four central protein layrs, assumed to be the 20S proteasome, with asymmetric V-shaped layrs of each end. These structural and functional characteristics of the spinach 26S proteasome showed marked similarity to those of the mammalian 26S proteasomes reported recently, suggesting that the 26S proteasome is widely distributed in eukaryotic cells and is of general importance for catalyzing the soluble energy-and ubiquitin-dependent proteolytic pathway.

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] K.Fujinami,T.Sato,H.Nakasawa: "Purification and Characterization of the 26s proteasome from spinach leaues" J.Biol.Chem,. 269. 25905-25910 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 中川弘毅: "植物からはじめて見いだされたプロテアリーム" 化学と生物. 32. 620-623 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Fujinami, K., Tanahashi, N., Tanaka, K., Ichihara, A., Cejka, Z., Baumeister, W., Miyawaki, M., Sato, T.and Nakagawa, H.: "Purification and characterization of the 26S proteasome from spinach leaves." J.Biol.Chem.269. 25905-25910 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Nakagawa, H.: "Proteasome form higher plant" Chemistry and Biology. 32. 620-623 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.Fujinami,T.Sato,H.Nakagawa: "Porification and characterization of the 26S Proteasome from spirach leaves" J.Biol.Chem. 269. 25905-25910 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 中川弘毅: "植物からはじめて見いだされたプロテアソーム" 化学と生物. 32. 620-623 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] M.Ozaki et al: "Purification and initial characterization of proteasome from the higher plant Spinacia oleracea" J.Biol.Chem. 267. 21678-21684 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] 藤波宏治 他: "高等植物におけるプロテアソームの精製と性質" 生化学. 65. 753 (1993)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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