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Crystal Structure Analysis of Bovine 20S Proteasome at High Resolution.

Research Project

Project/Area Number 07680723
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionHimeji Institute of Technology (1996-1997)
The University of Tokushima (1995)

Principal Investigator

MORIMOTO Yukio  Himeji Institute of Technology Associate Professor, 理学部, 助教授 (80200450)

Co-Investigator(Kenkyū-buntansha) TANAKA Keiji  The Tokyo Metropolitan Institute of Medical Science, 部長 (10108871)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1996: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1995: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsProteasome / Crystal Structure / Immuno-response / X線結晶解析 / X線構造解析 / 蛋白分解 / 細胞内蛋白分解
Research Abstract

Proteasome is a widery distributed protease in cellular organism, which plays an important role in a cell cycle, and proteolyze unneeded proteins to keep the cell being stable.
Eukaryotic proteasomes are multicatalytic proteinase complexes with a molecular weight of 750 kDa, containing, respectively, two copies of a hetero-heptamer of alpha-type subunits and one of beta-type subunits, (alpha1-7beta1-7)2. Proteasome was purified from bovine liver and crystallized into a hexagonal system with cell dimensions of a=b=121.83 (2) *, c=930.68 (6) *. A cylindrical particle size of 122 * diameter and 155* height was determined from the molecular packing in a unit cell. The srystal gave diffraction spots up to at least 4.4 * resolution, which was the minimum spacing of the camera used. The overall temperature factor of the enzyme was estimated to be in the range of 36.2A^2 to 25.8*^2. These results imply that the enzyme complex has a unique ordered structure comprising multisubunits with two types of hetero-heptamer. This ordered structure may facilitate highly organized cooperation of individual functions of subunits within the enzyme complex. And new crystal forms of the proteasome were obtained by changing the concentration of a precipitant, whose dimensions were 1/3 of the previous cell dimension.
An evaluation of the diffraction data from such crystal are now in progress.
In 1997, Prof.R.Huber in Max Plank Inst.clarified the tertiary structure of a yeast proteasome, and disscussed the mechanism of multi-proteolytic function at 2 A resolution. But, immuno-responsibility could not be described due to yeast-system. Therefore, proteasome and its activator are important target to show immuno-system in a Eukaryotic cell.

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] 森本 幸生: "X線解析によるプロテアーゼの構造解析" 蛋白質核酸酵素. 42. 2448-2453 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Morimoto, et.al.: "X-ray crystallographic studies on Bovine 20S Immuno-proteasome" Photon Factory Activity Report. 14. 75- (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 富杉 佳計 ほか: "プロテアソーム活性化因子PA28の精製と結晶化" 日本生物物理学予稿集. S11- (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Yabuuchi, et.al.: "Crystallization and Preliminary X-Ray Analysis of Salicylate Hydroxylase from Pseudomonas putida S-1." J.Biochem.119. 829-831 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Morimoto, et.al.: "Ordered Structure of the Crystallized Bovine 20S Proteasome." J.Biochem.117. 471-474 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 薮内 剛 ほか: "DIP2000/DENZOによるタンパク質結晶のX線回折強度データ収集" 技報. 8. 21-31 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Morimoto.: "Structural Studies on Proteases by Protein Crystallography" PROTEIN,NUCLEIC ACID AND ENZYME. 42. 2448-2453 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Tomisugi, et al.: "Purification and cryatallization of Proteasome activator, PA28." The proceeding of the Biophysical Society of Japan. S11 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Morimoto, et al.: "X-ray crystallographic studies on Bovine 20S Immuno-proteasome" Photon Factory Activity Report. 14. 75 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Yabuuchi, et al.: "Crystallization and Preliminary X-Ray Analysis of Salicylate Hydroxylase from Pseudomonas putida S-1." J.Biochem.119. 829-831 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Morimoto, et al.: "Ordered Structure of the Cryatallized Bovine 20S Proteasome." J.Biochem.117. 471-474 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Yabuuchi, et al.: "Intensity Data collection from protein srystals by using a DIP2000/DENZO system" GIHOU. 8. 21-31 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 森本幸生: "X線解析によるプロテアソームの構造解析" 蛋白質核酸酵素. 42. 2448-2453 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y.Morimoto, et.al.: "X-ray crystallographic studies on Bovine 20S Immuno-proteasome" Photon Factory Activity Report. 14. 75- (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] 富杉佳計ほか: "プロテアソーム活性化因子PA28の精製と結晶化" 日本生物物理学会予稿集. S11- (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Yabuuchi, et.al.: "Crystallization and Preliminary X-Ray Analysis of Salicylate Hydroxylase from Pseudomonas putida S-1" J.Biochem.119. 829-831 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y.Morimoto, et.al.: "Ordered Structure of the Crystallized Bovine 20S Proteasome" J.Biochem.117. 471-474 (1995)

    • Related Report
      1997 Annual Research Report
  • [Publications] 薮内剛ほか: "DIP2000/DENZOによるタンパク質結晶のX線回折強度データ収集" 技報. 8. 21-31 (1995)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y. Morimoto: "Ordered Structure of the Crystallized Bovine 20S Proteasome." J. Biochem.117. 471-474 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y.Morimoto: "Ordered Structure of the Crystallized Bovine 20S Proteasome" J.Biochem.117. 471-474 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 森本 幸生: "牛肝プロテアソームの結晶学的研究" 日本結晶学会 予稿集. 184-184 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] 水島恒裕: "ウシ肝臓プロテアソームの結晶化" 生化学. 66. 941-941 (1994)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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