Participation of Cytosolic Protein Phosphatase in Regulation of NADPH Oxidase in Polymorphonuclear Leukocytes

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概要

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• Calyculin A, a protein phosphatase inhibitor, enhanced phorbol 12-myristate 13-acetate (PMA)-induced superoxide anion (O^-_2) producuction and translocation of the cytosolic NADPH oxidase factor, p47phox, to the plasma membrane in guinea pig polymorphonuclear leukocytes (PMNs). When PMNs were treated with 1-(5-isoquinoline-sulfonyl)-3-methyl-piperazine (H-7), a protein kinase C (PKC) inhibitor, after exposure to PMA, inhibition of O^-_2 production and translocation of p47phox to the membrane fraction in PMA-stimulated PMNs were observed. When calyculin A was added to the PMA-stimulated PMNs after the addition of H-7,O^-_2 production was again observed, and translocation of p47phox to the membrane fraction also occurred. The activity of NADPH oxidase, the amount of p47phox and the level of phosphorylation of p47phox in the membrane fraction prepared form PMA-stimulated PMNs, were reduced by the addition of the cytosol fraction from unstimulated PMNs.These reductions were attenuated by calyculin A. These results indicated that the active from of NADPH oxidase in PMNs can be reconstituted after the active complex of the enzyme has disappeared once, and that one of the mechanisms of regulation of this enzyme activity involves the phosphorylation of p47phox in the cyotosol and dephosphorylation of phosphorylated p47phox in the NADPH oxidase complex by protein kinase and protein phosphatase, respectively.

• 公益社団法人日本薬学会の論文
• 1999-06-15

著者

• HAYAKAWA Takao

  National Institute of Health Sciences

• Shimohama S

  Kyoto Univ. Kyoto Jpn

• Shimohama Shun

  Department Of Neurology Faculty Of Medicine Kyoto University

• FUJIMOTO Sadaki

  Department of Environmental Biochemistry, Kyoto Pharmaceutical University

• 河上 奈保子

  京都薬大 衛生化学

• Hayakawa T

  Pharmaceutical Res. And Technol. Inst. Kinki Univ.

• Hayakawa Takao

  Division Of Biological Chemistry And Biologicals National Institute Of Health Sciences

• Kawakami Naomi

  Department Of Hospital Pharmacy Toyama Medical And Pharmaceutical University

• OKAMURA Naoki

  Department of Physiological Chemistry, Hiroshima University School of Medicine

• KAWAKAMI Naoko

  Department of Environmental Biochemistry, Kyoto Pharmaceutical University

• Okamura Naoki

  Department Of Physiological Chemistry Faculty Of Medicine Hiroshima University

• Okamura Naoki

  Department Of Physiological Chemistry Hiroshima University School Of Medicine

• Fujimoto Sadaki

  Department Of Environmental Biochemistry Kyoto Pharmaceutical University

• TAKEMASA Hiroaki

  Department of Applied Physics,Faculty of Science,Science University of Tokyo

• Takemasa Hiroaki

  Department Of Environmental Biochemistry Kyoto Pharmaceutical University

• Takemasa Hiroaki

  Department Of Applied Physics Faculty Of Science Science University Of Tokyo

• FUJIMOTO Sadaki

  Department of Biochemistry I Kyoto Pharmaceutical University:Department of III Kyoto Pharmaceutical University:Department of Pharmaceutical Chemistry II Kyoto Pharmaceutical University

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