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ROLE OF PHOSHOLIPASE D IN SIGNAL TRANSDUCTION AND ACTIVATION MECHANISM IN RABBIT PERITONEAL NEUTROPHILS

Research Project

Project/Area Number 04680186
Research Category

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

Allocation TypeSingle-year Grants
Research Field 代謝生物化学
Research InstitutionTOKYO INSTITUTE OF TECHNOLOGY (1993)
Gifu University (1992)

Principal Investigator

KANAHO Yasunori  TOKYO INSTITUTE OF TECHNOLOGY DEPARTMENT OF LIFE SCIENCE ASSOCIATE PROFESSOR, 生命理工学部, 助教授 (00214437)

Project Period (FY) 1992 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1993: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1992: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsPhospholipase D / Neutrophils / Signal transduction / Permeabilized cells / GTP-bnding protein / Calmodulin / Cytosol / Calcium / 好中球 / スタウロスポリン / 2、3ージホスホグリセリン酸 / GTP結合蛋白質 / 酵素放出反応 / 活性酸素
Research Abstract

It has recetly been reported that, in vaious types of mammalian cells, many agonists activates phospholipase D thrugh receptors on plasma membranes. The mechanism f the enzyme activation is nt yet been clarified. To investigate the activation mechanism f phospholipase D in rabbit peritoneal neutrophls, permeabilized neutrophils and cytosol prepared from rat brain were reconstituted and the phoshlipase D activation was investigated under various conditions.
[I] Phosholipase D ctivation in reconstituted system containing permeabilized neutrohils and cytosol - After prelabelng 1-omicron-alkyl-phoshatidylcholine in rabbit peritoneal neutrophils with [^3H]lyso platelet-activating factor at 37゚C for 2 hr, the cells were permeabilized with 1 U/ml of streptolysin Oat 37゚C for 20 mn to remove cytosol. The permeabilized cells were incubated in the presence of 1% ethanol under vaius conditions, then [^3H]phsphatidylethanl (PEt), an unambiguous product of phospholipase D, were determined. When cyto … More sol prepared from rat brain were reconstituted with permeabilized cells, GTPgammaS/Ca^<2+>/Mg-ATP significantly stimulted [^3H]PEt formation. The stimulation was not observed in the absence of cytosol. Since phospholipase D locates on plasma membranes, these results suggest that phospholipase D activation requires cytosolic factor(s).
[II] Phospholipase D activation factor : Since it is likely that a Ca^<2+>-dependent factor is, at least in part, involved in phospholipase D activation in rabbit peritoneal neutrophils it was investigated whether or not calmodulin (CaM) is involved in phospholipase D activation in the reconstituted system described above. CaM-free cytosol was prepared by flufenazine-immobilized affinity column chromatography. CaM bound to the column as eluted with EGTA slution. In permeabilized neutrophils reconstituted with CaM-free cytosol, GTPgammaS/Ca^<2+>/Mg-ATP stimulated phospholipase D to much less extent than in the reconstitute dsystem with cytosol. When CaM euted frm the column was added to the reconstituted system with CaM-free cytosol, phospholipase D activation by GTPgammaS/Ca^<2+>/Mg-ATP was maredly augmented. The urified CaM from bovine brain cytosl had the same effect with CaM eluted from the affinity column. From these results, it is suggested that CaM ia, at least in part, involved in phospholipase D activation as a cytosolic factor in rabbit peritoneal neutrophils. Less

Report

(3 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • Research Products

    (31 results)

All Other

All Publications (31 results)

  • [Publications] Hiroyuki Kanoh: "Requirement of adenosine 5'-triphosphate and Ca^<2+> for guanosine 5'-triphosphate-binding protein-mediated phospholipase D activation in rat pheochromocytoma PC12 cells" Neuros.Lett.151. 146-149 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Yasunori Kanaho: "The phosphatase inhibitor 2,3-diphosphoglycerate interferes with phospholipase D activation in rabbit peritoneal neutrophils" J.Biol.Chem.268. 12492-12497 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Yasunori Kanaho: "A protein kinase C inhibitor,staurosporine,activates phospholipase D via a pertussis toxin-sensitive GTP-binding protein in rabbit peritoneal neutrophils" J.Biol.Chem.267. 23554-23559 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Akira Nishida: "Characterization of phospholipase D in a cell-free system of cultured cells derived from rat frontal cortex" Brain Res.595. 12-16 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Hiroyuki Kanoh: "Pertussis toxin-insensitive G protein mediates carbachol activation of phospholipase D in rat pheochromocytoma PC12 cells" J.Neurochem.59. 1786-1794 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Yasunori Kanaho: "Calcium rather than protein kinase C is the major factor to activate phospholipase D in fMLP-stimulated rabbit peritoneal neutrophils" J.Immunol.149. 622-628 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 金保 安則: "実験医学" 羊土社, 7 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 金保 安則: "神経研究の進歩" 医学書院, 7 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Hiroyuki Kanoh: "Requirement of adenosine5'-triosphate and Ca^<2+> for guanosine 5'-triphosphate-binding proteinmediated phospholipase D activation in rat pheochroocytoma PC12 cells" Neuros.Lett. 151. 146-149 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Yasunori Kanaho: "The phosphatase inhibitor 2,3-diphoshoglycerate interferes with phospholipase D activation in rabbit peritoneal neutrophils" J.Biol.Chem. 268. 12492-12497 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Yasunori Kanaho: "A protein kinase C inhibitor, staurosporine, activates phospholipase D via a pertussis toxin-sensitive GTP-binding protein in rabbit peritoneal neutrophils" J.Biol.Chem. 267. 23554-23559 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Akira Nishida: "Characterizaation of phospholipase D in a cell-free system f cutured cells derived from rat frontal cortex" Brain Res. 595. 12-16 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Hiroyuki Kanoh: "Pertssis toxin-insensitive G protein mediates carbachol activation of phospholipase D in rat pheochromocytoma PC12 cells" J.Neurochem. 59. 1786-1794 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Yasunori Kanaho: "Calcium rather than protein kinase C is the major factor to activate phospolipase D in fMLP-stimulated rabbit peritoneal neutrophils." J.Immunol. 149. 622-628 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Yasunori Kanaho: "The mechanism of phospholipase D activation" Exprimental Medicine. 11. 31-37 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Yasunori Kanaho: "Membrane phospholipid metabolism in signal transduction" Advances in Neurological Sciences. 36. 466-475 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Hiroyuki Kanoh: "Requirement of adenosine5′-triphosphate and Ca^<2+> for guanosine 5′-triphosphate-binding protein-mediated phospholipase D activation in rat pheochromocytoma PC12 cells" Neuros.Lett.151. 146-149 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Yasunori Kanaho: "The phosphatase inhibitor 2,3-diphosphoglycerate interferes with phospholipase D activation in rabbit peritoneal neutrophils" J.Biol.Chem.268. 12492-12497 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Yasunori Kanaho: "A protein kinase C inhibitor,staurosporine,activates phospholipase D via a pertussis toxin-sensitive GTP-binding protein in rabbit peritoneal neutrophils" J.Biol.Chem.267. 23554-23559 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Akira Nishida: "Characterizaation of phospholipase D in a cell-free system of cutured cells derived from rat frontal cortex" Brain Res.595. 12-16 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Hiroyuki Kanoh: "Pertussis toxin-insensitive G protein mediates carbachol activation of phospholipase D in rat pheochromocytoma PC12 cells" J.Neurochem.59. 1786-1794 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Yasunori Kanaho: "Calcium rather than protein kinase C is the major factor to activate phospholipase D in fMLP-stimulated rabbit peritoneal neutrophils" J.Immunol.149. 622-628 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] 金保 安則: "実験医学" 羊土社, 7 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 金保 安則: "神経研究の進歩" 医学書院, 7 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Y.Kanaho et al.: "Calcium rather than protein kinase C is the major to activate phospholipase D in fMLP-stimulated rabbit peritoneal neutrophils.Possible involvement of calmodulin/myosin L chain kinase pathway." J.Immunol.149. 622-628 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] H.Kanoh et al.: "Pertusis toxin-insensitive G protein mediates carbachol activation of phospholipase D in rat pheochromocytoma PC 12 cells." J.Neurochem.59. 1786-1794 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] A.Nishida et al.: "Characterization of phospholipase D in a cell-free system of cultured cells derived from rat frontal cortex." Brain Res.595. 12-16 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Y.Kanaho et al.: "A protein kinase C inhibitor,staurosporine,activates phospholipase D via a pertusis toxin-sensitve GTP-binding protein in rabbit peritoneal neutrophils." J.Biol.Chem.267. 23554-23559 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] H.Kanoh et al.: "Requirement of ATP and Ca^<2+> for GTP-binding protein-mediated phospholipase D activation in rat pheochromocytoma PC 12 cells." Neurosci.Lett.(in press).

    • Related Report
      1992 Annual Research Report
  • [Publications] Y.Kanaho et al.: "The phosphatase inhibitor 2,3-diphosphoglycerate interferes with phospholipase D activation in rabbit peritoneal neutrophils." J.Biol.Chem.(in press).

    • Related Report
      1992 Annual Research Report
  • [Publications] 金保 安則、ほか: "神経研究の進歩:神経系の細胞骨格・細胞内情報伝達系" 医学書院, 532 (1992)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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