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1991 Fiscal Year Final Research Report Summary

Production of active oxygen metabolites in polymorphonuclear leukocytes and regulatory mechanism for the production

Research Project

Project/Area Number 01480492
Research Category

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

Allocation TypeSingle-year Grants
Research Field Biological pharmacy
Research InstitutionHiroshima University

Principal Investigator

ISHIBASHI Sadahiko  Hiroshima University, School of Medicine, Professor, 医学部, 教授 (90012616)

Co-Investigator(Kenkyū-buntansha) OKUMURA Naoki  Hiroshima University, School of Medicine, Research associate, 医学部, 助手 (30144827)
Project Period (FY) 1989 – 1991
KeywordsActive Oxygen / Superoxide Anion / Polymorphonuclear Leukocytes / NADPH Oxidase / Protein Kinase C / Cell Membrane / Enzyme Regulation
Research Abstract

This research project was planned to characterized NADPH oxidase as well as to elucidate the mechanism for the activation of this enzyme which is dormant in the absence of stimulation. Guinea pig peritoneal polymorphonuclear leukocytes were used as the experimental material.
We previously found that cytosolic 46kDa protein was phosphorylated in parallel with 0^2_ production and protein kinase C (PKC) was responsible for the phosphorylation. In the present study, translocation of this protein was found from the cytosol to the cell membranes. It was assumed that active NADPH oxidase complex was formed by the association of the cytosolic protein (s) and membranous proteins, flavoprotein, cytochrome b, etc., and that the translocation was responsible for the activation. As the significance of the phosphorylation by PKC, we proposed from the similar effect of arachidonate and SDS that the phosphorylation worked through introduction of negative charge to the cytosolic protein. In the later st … More age of the present study, we recognized that the 46kDa protein corresponded probably to one
of the cytosolic activating factors reported for human leukocytes.
As another evidence for the active complex formation, we found that performed NADPH oxidase complex was greatly stabilized by the treatment of the leukocytes with glutaraldehyde at very low concentration. On the contrary, the pretreatment with glutaraldehyde canceled the activation of NADPH oxidase.
Next, we examined preceding (so-called priming) condition for O^2_ production. As compared with isotonic condition, the stimulation under hypotonic condition caused much increase in the production. Treatment of the leukocytes with cytochalasin had similar effect. On the other hand, change in depolarization pattern was observed in the stimulated leukocytes in accordance with the degree of the change in 0^2_ production. These findings indicate that membrane potential and /or structure is involved in the production through providing the priming ondition in the membranes prior to the active complex formation. Less

  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] Masaki Ozawa: "Synergism between Protein Kinase C Activatior and Fatty Acids in Stimulating Superoxide Anion Production in Guinea Pig Polymorphonuclear Leukocytes" Arch.Biochem.Biophys.273. 491-496 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toshiaki Ohtsuka: "Stimulatory Effects of a Short Chain Phosphatidate on Superoxide Anion Production in Guinea Pig Polymorphonuclear Leukocytes" J.Biochem.106. 259-263 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toshiaki Ohtsuka: "Synergism between Platelet-Activating Factor and Diacylglycerol in the Induction of Superoxide Anion Production in Guine Pig Polymorphonuclear Leukocytes" Arch.Biochem.Biophys.279. 21-24 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Teruaki Katayama: "Effect of Glutaraldehyde on NADPH Oxidase System of Guinea Pig Polymorphonuclear Leukocytes" Arch.Biochem.Biophys.278. 431-436 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toshiaki Ohtsuka: "Involvement of Membrane Charges in Constituting the Active Form of NADPH Oxidase in Guinea Pig Polymorphonuclear Leukocytes" Arch.Biochem.Biophys.280. 74-79 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kyoko Wakamuta: "Mechanism for the inhibitory Effect of a Seleno-Organic Compound,Ebselen,and Its Analogues on Superoxide Anion Production in Guinea Pig Polymorphonuclear Leukocytes" J.Pharmacobio-Dyn.13. 421-425 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toshiaki Ohtsuka: "Translocation of the 46kDa Protein(S) in Response to Activation of NADPH Oxidase in Guinea Pig Polymorphonuclear Leukocytes" J.Biochem.108. 169-174 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toshiaki Ohtsuka: "A Diacylglycerol Kinase Inhibitor,R 59 022,Potentiates Superoxide Anion Production and 46-kDa Protein Phosphorylation in Guinea Pig Polymorphonuclear Leukocytes" J.Biol.Chem.265. 15418-15423 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 石橋 貞彦: "好中球の活性酸素生成と蛋白質リン酸化反応" J.Act.Oxyg.Free Rad.1. 171-182 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masanori Hiura: "Stimulation of Superoxide Anion Production in Guinea Pig Polymorphonuclear Leukocytes by Hypotonic Conditions in Combiantion with Protein Kinase C Activatiors" Arch.Biochem.Biophys.291. 31-37 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sadahiko Ishibashi: "Mechanism for production of active oxygen metabolites on plasma membranes of neutrophil" Membrane. 16. 131-140 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sadahiko Ishibashi(共同執筆): "Physiology and pharmacology of transmembrane signalling" Excerpta Medica, 404 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 石橋 貞彦(共同執筆): "白血球と生体防御" 講談社, 272 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ozawa, M., Ohtsuka, T., Okamura, N., Ishibashi, S: "Synergism between Protein Kinase C Activator and Fatty Acides in Stimulating Superoxide Anion Production in Guinea Pig Polymorphonuclear Leukocytes" Arch. Biochem. Biophys.273. 491-496 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ohtsuka, T., Ozawa, M., Okamura, N., Ishibashi, S.: "Stimulatory Effects of a Short Chain Phosphorylation on Superoxide Anion Production in Guinea Pig Polymorphonuclear Leukocytes" J. Biochem.106. 259-263 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Katayama, Y., Ohtsuka, T., Wakamura, K., Yoshida, K., Okamura, N., Isibashi, S.: "Effect of Glutaraldehyde on NADPH Oxidase System of Guinea Pig Polymerphonuclear Leukocytes" Arch. Biochem. Biophy.278. 431-436 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ohtsuka, T., Ozawa, M., Okamura, N., Ishibashi, S.: "Synergism between Platelet-activating Factor and Diacylglycerol in the Induction of Super-oxide Anion Production in Guinea Pig Polymorphonuclear Leukocytes" Arch. Biochem. Biophy.279. 21-24 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ohtsuka, T., Hiura, M., Ozawa. M., Okamura, N., Nakamura, M., Ishibashi, S.: "Involvement of Membrane Charges in Constituting the Active Form of NADPH Oxidase in Guinea Pig Polymorphonuclear Leukocytes" Arch. Biochem. Biophys.280. 74-79 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Wakamura, K., Ohtsuka, T., Okamura, N., Ishibashi, S., Masayasu, H.: "Mechanism for the Inhibitory Effect of a Seleno-Organic Compound, Ebselen, and Its Analogues on Superoxide Anion Production in Guinea Pig Polymorphonuclear Leukocytes" J. Pharmacobio-Dyn.13. 421-425 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ohtsuka, T., Nakamura, M., Hiura, M., Yoshida, K., Okamura, N., Ishibashi, S.: "Translocation of the 46kDa Protein (s) in Response to Activation of NADPH oxidase in Guinea Pig Polymorphonuclear Leukocytes" J. Biochem.108. 19-174 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ohtsuka, T., Hiura, M., Yoshida, K., Okamura, N., Ishibashi, S.: "A Diacylglycerol Kinase Inhibitor, R 59 022, Potentiates Superoxide Anion Production and 46-kDa Protein Phosphorylation in Guinea Pig Polymorphonuclear Leukocytes" J. Biol. Chem.265. 15418-15423 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hiura, M., Ozawa, M., Ohtsuka, T., Takesue, H., Yamaguchi, M., Ikamura, N., Ishibashi, S.: "Stimulation of Superoxide Anion Production in Guinea Pig Polymorphonuclear Leukocytes by Hypotonic Condition in Combination with Protein Kinase C Activators" Arch. Biochem. Biophys.291. 31-37 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ishibashi, S., Okamura, N.: "Mechanism for production of active oxygen metabolites on plasma membranes of neutrophils" Membrane. 16. 131-140 (1991)

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

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Published: 1993-03-16  

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