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Analysis for the Substrate Strucure and Structure Responsible for Substrate-recognition by Mitochondrial Processing Peptidase

Research Project

Project/Area Number 07680692
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Functional biochemistry
Research InstitutionKYUSHU UNIVERSITY

Principal Investigator

OGISHIMA Tadashi  Kyushu University, Chemistry, Associate Professor, 理学部, 助教授 (70177153)

Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1996: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1995: ¥1,200,000 (Direct Cost: ¥1,200,000)
KeywordsProcessing Peptidase / Protease / Substrate-recognition / Photoaffinity Labelling / プロセシングプロテアーゼ / ペプチド基質 / ミトコンドリアタンパク質前駆体 / 延長ペプチド / 分子認識 / フォトアフィニティーラベル
Research Abstract

I developed fluorogenic peptides, which were applicable for a rapid and real-time measurement of the acitivity of mitochondrial processing peptidase (MPP). Elements in the substrate required for processing by MPP were essentially the same as obtained from the previous kinetical study using synthetic peptides modeled on the extension peptides of rat malate dehydrogenase (MDH). Primary recognition elements were the distal basic amino acid and proximal arginine residues, and the amino acid at P1' position. The portion between the distal and proximal amino acids proved to be flexible for the efficient processing. I found that MPP recognizes the proximal arginine via two sets of hydrogen-bonding and one ionic bonding.
From a mutation of MPP,beta-subunit was determined to be the catalytic subunit. I succeeded in expressing and purifying recombinant enzyme of both subuits. Photoaffinity labelling of the subuints demonstrated that the substrate binds to both subuints in the manner that different portions of the substate were in contact with different subunits.

Report

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

    (9 results)

All Other

All Publications (9 results)

  • [Publications] Song,M-C.: "Role of the Basic Amino Acids in the Cleavage of Synthetic Peptide Substrates by Mitochondrial Processing Peptidase" J.Biochem.120. 1163-1166 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ogishima,T.: "Analysis of Elements in the Substrate Reqired for Processing by Mitochondrial Processing Peptidase." J.Biol.Chem.270. 30322-30326 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kitada,S.: "A Putative Metal-binding Slte in th b Subunit of Rat Mtochondrial Processing Peptidase is Essential for Its Catalytic Activity" J.Biochem.117. 1148-1150 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Song, M-C., Shimokata, K., Kitada, S., Ogishima, T., & Ito, A.: "Role of the Basic Amino Acids in the Cleavage of Synthetic Peptide Substrates by Mitochondrial Processing Peptidase" J.Biochem.120. 1163-1166 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ogishima, T., Niidome, Shimokata, K., Kitada, S., & Ito, A.: "Analysis of Elements in the Substrate Reqired for Processing by Mitochondrial Processing Peptidase" J.Biol.Chem.270. 30322-30326 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kitada, S., Shimokata, K., Niidome, T., Ogishima, T., & Ito, A.: "A Putative Metal-binding Site in the beta Subunit of Rat Mtochondrial Processing Peptidase is Essential for Its Catalytic Activity" J.Biochem.117. 1148-1150 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 宋明哲: "Role of Basic Amino Acids in the Cleavage of Synthetic Peptide Substrates by Mitochondrial Processing Peptidase" J.Biochem.120(6). 1163-1166 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 荻島正: "Analysis of Elements in the Substrate Required for Processing by Mitochondrial Processing Peptidase" Journal of Biological Chemistry. 270. 30332-30326 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 北田栄: "A Putative Metal-Binding Site in the β-Subunit of Rat Mitochondrial Processing Peptidase is Essential for Its Catalytic Activity" Journal of Biochemistry. 117. 1148-1150 (1995)

    • Related Report
      1995 Annual Research Report

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

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