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Study of Substrate-recognition Mechanism of Zn-metalloprotease

Research Project

Project/Area Number 05680580
Research Category

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

Allocation TypeSingle-year Grants
Research Field Biophysics
Research InstitutionUniversity of Tokyo (1994)
Osaka University (1993)

Principal Investigator

HARADA Shigeharu  Univ.of Tokyo, Dept.of Pharmaceutical Technochemistry, 薬学部, 助教授 (80156504)

Co-Investigator(Kenkyū-buntansha) 甲斐 泰  大阪大学, 工学部, 教授 (40029236)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1994: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1993: ¥1,100,000 (Direct Cost: ¥1,100,000)
Keywordsprotease / metalloprotease / X-ray crystal structure analysis / Streptomyces caespitosus / Streptomayces caespitosus
Research Abstract

The three-demensional structure and amino acid sequence of a zinc metalloprotease produced by Streptomyces caespitosus (SCNP) have been determined in order to clarify the relationship between structure and function of SCNP.The amino-acid sequence has been determined by Edman degradation. SCNP consists of a single polypeptide chain of 132 amino acid residues with one disulfide bond between residues 99 and 112. The deduced amino acid sequence indicated that it is much shorter than other ones from metalloproteases previously reported. Although a zinc-binding motif, HEXXH,found at the active sites of most metalloproteases, was found in the sequence, SCNP did not share overall significant similarity to the sequences of other zinc metalloproteases. The three-demensional structure of SCNP has been determined by X-ray crystal structure analysis and refined to R-factor of 0.18 (2.0A resolution). A five stranded beta-sheet and three alpha-helices are found in the structure. The zinc-binding moti … More f (H83-E-T-G-H87) is located on the second alpha-helix from N-terminal. About 40 amino acid sequences of zinc-containing metalloproteases have been determined so far and classified into five distinct families according to the sequence homology : thermolysin, astacin, serratia, matrixin and snake venom. Crystal structures available for six enzymes have revealed that two histidine residues located in the the sequence of HEXXH are the first two zinc ligands. Gly observed as the 8th residue from the first His in the motif, which is conserved in all but thermolysin family, has been reported to be important for structural reason, becaluse this residue allows the 11th His to be the third zinc ligand by terminating the second alpha-helix and bending a main chain sharply. SCNP also has Gly at this position. However, the residue corresponding to the 11th His is Asp. The three-dimensional structure of SCNP revealed that this Asp is the third zinc lingand. It is deduced that SCNP may represent a new subfamily of zinc-containing metalloprotease, with respect to both the new type of ligand organization for Zn and a very small molecular size distinct from other known metalloproteases in the five families. Less

Report

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

    (9 results)

All Other

All Publications (9 results)

  • [Publications] Shigeharu Harada: "Crystallization and Preliminary X-ray Studies of a Protease from Pseudomonas aeruginosa" J. Mol. Biol.230. 1315-1316 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] G.Kurisu: "Crystal Structure of Neutral Protease from Streptomyces caespitosus" Acta Cryst. A. A49. 74 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 原田繁春: "Streptomyces caespitosusが産出する蛋白質分解酵素の構造" 生産と技術. Vol.46. 55-57

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shigeharu Harada, Akinobu Nagara, Genji Kurisu & Yasushi Kai: "Crystallization and Preliminary X-ray Studies of a Protease from Pseudomonas aeruginosa" J.Mol.Biol. Vol.230. 1315-1316 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] G.Kurisu, A.Nagara, S.Harada, Y.Kai & N.Kasai: "Crystal Structure of Neutral Protease from Streptomyces caespitosus" Acta Cryst.A. Vol.A49. 74 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shigeharu Harada: "Structure of a protease produced by Streptomyces caespitosus" The Production & Technique. Vol.46. 55-57 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shigeharu Harada: "Crystallization and Preliminary X-ray Studies of a Protease from Pseudomonas aeruginosa" J.Mol.Biol.230. 1315-1316 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] G.Kurisu: "Crystal Structure of Neutral Protease from Streptomyces caespitosus" Acta Cryst.A. A49. 74 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] 原田繁春: "Streptomayces caespitosusが産出する蛋白質分解酵素の構造" 生産と技術. Vol.46. 55-57 (1994)

    • Related Report
      1994 Annual Research Report

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

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