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1997 Fiscal Year Final Research Report Summary

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
KeywordsProteasome / Crystal Structure / Immuno-response
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.

  • Research Products

    (12 results)

All Other

All Publications (12 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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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

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Published: 1999-03-16  

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