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Studies on the separation of phospholipase A_2 preferred the arachidonoyl species of phospholipids

Research Project

Project/Area Number 62580135
Research Category

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

Allocation TypeSingle-year Grants
Research Field 物質生物化学
Research InstitutionFaculty of Pharmaceutical Sciences, Teikyo University

Principal Investigator

NAKAGAWA Yasuhito  Fac. Pharm. Sci., Teikyo University, Lecturer, 薬学部, 講師 (00119603)

Project Period (FY) 1987 – 1988
Project Status Completed (Fiscal Year 1988)
Budget Amount *help
¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1988: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1987: ¥1,100,000 (Direct Cost: ¥1,100,000)
KeywordsMacrophage / Arachidonic Acid / Phospholipase / Calcium / カルモデュリン / リン脂質分子種
Research Abstract

The effects of calmodulin antagonists on the amount of free fatty acid produced by alveolar macrophages exposed to opsonized zymosan were determined. Opsonized zymosan-induced free arachidonic acid (20:4) production was dramatically suppressed in the presence of W-7 and trifluoperazine without an effect on the production of other fatty acids. Calmodulin antagonists appeared to inhibit phospholipase A_2, since W-7 abolished the liberation of 20:4 from choline glycerophospholipids (CGP) and inositol glycerophospholipids (IGP$. After the challenge with opsonized zymosan, the 20:4 liberation caused by opsonized zymosan was significantly reduced in the absence of extracellular Ca^<2+>, indicating that the release of arachidonic acid required the influx of the extracelluler Ca^<2+> into cell. On the other hand, oleic acid and linoleic acid liberation from alveolar macrophages exposed to opsonized zymosan was not inhibited after the depletion of extracellular Ca^<2+>.
Most of phospholipase A_2 activity was present in the cytosol fraction of alveolar macrophages, representing 70% of the total activity. Calcium enhanced the hydrolysis of 16:0-20:4 GPC with cytosolic phospholipase A_2, although the phdrolysis of 16:0-18:1 and 16:0-18:2 species of CGP was not changed by calcium.
The purification of cytosolic phospholipase A_2 has been performed by using the hydrophordic column, ion exchange column and gel filtration column. However, phospholipase A_2 which selectively hydrolyzed the arechidonoyl species of phospholipids failed to be isolated.
The present studies indicate that phospholipase A_2 in rabbit alveolar macrophages is controlled by Ca^<2+> and calmodulin, and preferentially hydrolyzes arachidonic acid from phospholipids.

Report

(3 results)
  • 1988 Annual Research Report   Final Research Report Summary
  • 1987 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Nakagawa,Y.: Biochem.Biophys.Res.Commun.156. 947-953 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Nakagawa,Y.: Arch.Biochem.Biophys.268. inpress (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Nakagawa,Y.: J.Choromatogr.in press.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Rustow,B.: Biochem.Biophys.Acta. 961. 364-369 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Rustow,B.: Biochem.J.254. 67-71 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Naito,M.: J.Biochem.102. 155-162 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Y. Nakagawa; K. Waku: "Selective inhibition of free arachidonic acid production in activated alveolar macrophages by calmodulin antagonists." Biochem. Biophys. Res. Commun.156. 947-953 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Y. Nakagawa; B. R stow; H. Rabe; D. Kunze; K. Waku: "The de novo synthesis of molecular species of phosphatidylinositol from endogenously labeled CDP diacylglycerol in alveolar macrophage microsomes." Arch. Biochem. Biophys.268. (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Y. Nakagawa; K. Waku: "Separation of the molecular species of diacyl and alkenylacyl subclasses of methyl ester of dinitrophenyl ethanolamine glycerophospholipid by high-performance liquid chromatography." J. Chromatogr.(1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] B. R stow; Y. Nakagawa; H. Rabe; G. Reichmann; D. Kunze; K. Waku: "Comparison of the HPLC-separated species patterns of phosphatidic acid, CDP-diacylglycerol and diacylglycerol synthesized de novo in rat liver microsomes (a new method)." Biochim. Biophys. Acta. 961. 364-369 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] B. R stow; Y. Nakagawa; H. Rabe; K. Waku; D. Kunze: "Species pattern of phosphatidylinositol from lung surfactant and a comparison of the species pattern of phosphatidylinositol and phosphatidylglycerol synthesized de novo in lung microsomal fractions." Biochem. J.254. 67-71 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Nakagawa,Y.: Biochem.Biophys.Res.Commun.156. 947-953 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Nakagawa,Y.: Arch.Biochem.Biophys.268. inthepress (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] Nakagawa,Y.: J.Chromatogr.

    • Related Report
      1988 Annual Research Report

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Published: 1987-04-01   Modified: 2016-04-21  

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