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1999 Fiscal Year Final Research Report Summary

Study on Mechanisms of ATP Synthesis in Single Mitochondria

Research Project

Project/Area Number 10680627
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionTokyo University of Agriculture and Technology

Principal Investigator

OHTA Yoshihiro  Tokyo University of Agriculture and Technology, Faculty of Engineering Associate Proffessor, 工学部, 講師 (10223843)

Project Period (FY) 1998 – 1999
Keywordsmitochondria / ATP / fluorescence microscopy / imaging / permeability transition / membrane potential / NADH / flavin
Research Abstract

The mitochondrial activities related to ATP production were measured in single mitochondria with fluorescence microscopy. So far, studies on mitochondria have been performed with mitochondrial suspension. Therefore, it has been impossible to detect asynchronous phenomena like the opening and the closing of a channel. In the present study, we developed the technique to measure the activity of single mitochondria to detect the phenomena that has not been detected with mitochondria suspension.
To observe single mitochondria with fluorescence microscopy, mitochondria were adsorbed on the cover slip coated with polyphenolic protein. Since the adsorption did not decrease the ATP production rate in mitochondria, this method for adsorption was elucidated to be useful for the measurements of mitochondrial activity. Upon addition of malate as a respiratory substrate, the time-courses of NADH formation and Flavin reduction in single mitochondria was similar to the results obtained for mitochondrial suspension. On the other hand, mitochondria showed the generation and disruption of the membrane potential due to the opening and the closing of the permeability transition pore. The opening and the closing depends on the redox state of the electron transport chain.

  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] Yamazaki et al.: "Calcium Ion as a Second Messenger for O-Nitrophenyl Sulfenyl - Adrenocorticotropin (NPS-ACTH) and ACTH in Bovine Adrenol Steroidogenesis"Endocrinology. 139. 4765-4771 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Suzuki-Tomii et al.: "Comparative Experiments of Protein Denaturation for Studying Structural Difference of Homologous Membrane Proteins : P450scc and P450_<11β>"Jpn.J.Appl.Phys.. 38. 298-301 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamada et al.: "Dynamic Mobility of Genetically Expressed Fusion Protein between Cytochrome P450(A) and NADPH-Cytochrome P450 reductase in Yeast Microsomes."Biochemistry. 38. 9465-9470 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 渡辺功一他: "蛍光顕微鏡による単離ミトコンドリアの観察方法"生物物理. 224. 258-260 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hoolahan et al.: "Molecular Steroidogenesis"Calcium Ion Enhances the Production of NAD(P)H and Pregnenolone in Bovine Adrehocortical Mitochondria. 442 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamazaki et al.: ""Calcium Ion as a Second Messenger for O-Nitrophenyl Sulfenyl-Adreno corticotropin(NPS-ACTH) AND ACTH in Bovine Adrenal Steroidogenesis""Endocrinology. 139. 4765-4771 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Suzuki-Tomii et al.: ""Comparative Experiments of Protein Denaturation for Studying Structural Difference of Homologous Membrane Proteins : P450scc and P45011β"Jpn. J.Appl. Phys.. 38. 289-301 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamada et al.: ""Dynamic Mobility of Genetically Expressed Fusion Protein between Cytochrome P450IAI and NADPH-Cytochrome P450 Reductase in Yeast Microsomes""Biochemistry. 38. 9465-9470 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hoolahon et ai.: ""Calcium Ion Enhances the Production of NAD(P)H and Pregnenolone in Bovine Adrenocortical Mitochondria""Moleculor Steroidogenesis. 442 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Watanabe et al.: ""Measurements of Single Mitochondria with Fluorescence Microscopy"(in Japanese)"Seibutsu-Butsuri. 224. 258-260 (1999)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2002-03-26  

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