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Development of a fast scanning IR spectrophotmeter and its application to oxygenase systems.

Research Project

Project/Area Number 59880012
Research Category

Grant-in-Aid for Developmental Scientific Research

Allocation TypeSingle-year Grants
Research Field 物質生物化学
Research InstitutionKeio University

Principal Investigator

ISHIMURA Yuzuru  慶応義塾大学, 医, 教授 (40025599)

Project Period (FY) 1984 – 1985
Project Status Completed (Fiscal Year 1985)
Budget Amount *help
¥9,700,000 (Direct Cost: ¥9,700,000)
Fiscal Year 1985: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1984: ¥7,800,000 (Direct Cost: ¥7,800,000)
KeywordsMechanisms of oxygen activation / biological oxidation / infrared spectrum / rapid scan infrared spectroophotometer / 二原子酸素伸縮振動 / ストップドフロー赤外計
Research Abstract

Aerobic organisms utilize molecular oxygen ( <O_2> ) in many ways. Such oxygen utilizing reactions include various oxidase and oxygenase reactions, and superoxide-producing reactions. Under physiologic conditions, however, molecular oxygen is mainly in an inactive triplet groud state which usaully requires an "activation" to react with singlet organic molecules. Thus, the mechanisms for activation of molecular oxygen has been one of the main themr in biochrmistry. In the present study, we developed a new type of IR spectrophotometer which enables us to measure O-O stretching frequency around 1100 <cm^(-1)> of molecular oxygen bound to iron in heme-containing oxygenases and oxidases. Since such oxygen complexes are all unstable reaction intermediates, it requires fast scanning together with high sensitivity, which are not fullfilled by any dispersive and/or FT type of IR spectrophotometer. For this purpose, we succesfully developed a new type of machine by employing MCT, high frequency chopper and high speed scanning device with special soft wear systems. The spectrophotmeter system made us possible to measure the O-O stretching of oxygen in oxy-CoMb (4 mM) within 7 minutes at 10゜C. In preliminary experiments, we also succeeded in measuring the O-O stretching of oxy-P450cam which was stabilized by utilizing the diacetylheme-substituted enzyme.

Report

(1 results)
  • 1985 Final Research Report Summary
  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] 第9回国際ラマン分光会議(東京)予稿集. 492 (1984)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1985 Final Research Report Summary
  • [Publications] J.Biol.Chem.260-No.3. (1985)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1985 Final Research Report Summary
  • [Publications] J.Biol.Chem.260-No.4. (1985)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1985 Final Research Report Summary
  • [Publications] J.Biol.Chem.261. (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1985 Final Research Report Summary
  • [Publications] Proceedings of the 7th International Congress on Raman spectroscopy. p.492-493 (1984)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1985 Final Research Report Summary
  • [Publications] J. Biol. Chem.260 - No.3. (1985)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1985 Final Research Report Summary
  • [Publications] J. Biol. Chem.260 - No.4. (1985)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1985 Final Research Report Summary
  • [Publications] J. Biol. Chem.261. (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1985 Final Research Report Summary

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Published: 1987-03-31   Modified: 2016-04-21  

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