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Intracellular Na^+ Influx by Extracellular Stimuli and Signal Transduction

Research Project

Project/Area Number 62570126
Research Category

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

Allocation TypeSingle-year Grants
Research Field Pathological medical chemistry
Research InstitutionFukui Medical School

Principal Investigator

HASHIMOTO Eikichi  Fukui Medical School, Associate Professor, 医学部, 助教授 (20116239)

Co-Investigator(Kenkyū-buntansha) SAKANOUE Youichiro  Fukui Medical School, Research Associate, 医学部, 助手 (00187030)
Project Period (FY) 1987 – 1988
Project Status Completed (Fiscal Year 1988)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1988: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1987: ¥1,100,000 (Direct Cost: ¥1,100,000)
KeywordsCell Proliferation / Na^+ / H^+ Exchanger / Ca^<2+>-Phospholipid-Dependent Protein Kinase (C-Kinase) / S6 Protein / Ion Signalling / Protein Phosphorylation / Limited proteolysis / プロテアーゼ / Ca^<2+>-燐脂質依存性 プロテインキナーゼ(Cキナーゼ) / 限定分解反応 / 細胞内Na^<+ >流入 / 細胞増殖因子 / 増殖因子 / Ca^<2+>-リン脂質依存性プロテインキナーゼ(Cキナーゼ) / 蛋白質リン酸化反応 / 制限消化
Research Abstract

In an attempt to understand the role of Na^+-influx and cytoplasmic alkalinization induced by various growth factors and tumor-promoting phorbol esters, the effect of ionic strength and pH was examined on activation of membrane-bound protein kinases. During these experiments, Ca^<2+> and phospholipid-dependent protein kinase (protein kinase C) was shown to be activated by trypsin-like protease in an ionic strength (140-210 mM NACL) and pH (pH7-8)-dependent manner. This reaction required Ca^<2+> and phospholipid and protein kinase C with molecular mass of 80,000 was converted to the active fragment of this enzyme (protein kinase M) with molecular mass of 45,000. This kind of proteolytic activation of protein kinase C was also demonstrated in regenerating rat liver 5 h after partial hepatectomy. The properties of this activated kinase detected in regenerating liver was closelysimilar with those of protein kinase M and this enzyme activity was detected with rebosomal proteins and histones. It is well-established that the phosphorylation of S6 protein in 40s rebosomal subunit is observed in various cellular systems stimulated by many growth factors and phorbol esters. In order to elucidate the function of protein kinase M, the phosphorylated sites of S6 protein by this enzyme was determaned using synthetic peptide analogue of thie protein. By the analysis of the phoshporylated peptide, Ser-236, Ser-240 and Ser-242 were identified as major phosphorylated sites in S6 protein. These sites corresponded to some of the multiple serine residues whose phosphroylation were confirmed by in vivo system. These accumulated evidences strongly suggest that protein kinase M proteolytically produced from protein kinase C after stimulation of Na^+/H^+ exchanger may have an important role at the early stage of cell proliferation.

Report

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

Research Products

(25 results)

All Other

All Publications (25 results)

  • [Publications] Akira Takeda: FEBS Letters. 210. 169-172 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Keiko Sakai: Biochemistry International. 14. 63-70 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Keiko Mizuta: Biochem.Biophys.Res.Commun.146. 239-246 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Youichiro Sakanoue: European Journal of Biochemistry. 168. 669-677 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Youichiro Sakanoue: Biochem.Biophys.Res.Commun.150. 1176-1184 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Kaoru Nishiyama: Biochemistry International. 17. 51-58 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Eikichi Hashimoto: Journal of Biochemistry. 104. 934-938 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Masahiko Hoshijima: Biochem.Biophys.Res.Commun.157. 851-860 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Akira Takeda: "Phosphorylation of liver gap junction protein by protein kinase C." FEBS Letters. 210. 169-172 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Keiko Sakai: "Non-requirement of calcium on protamine phosphorylation by calcium-activated, phospholipid-dependent protein kinase." Biochemistry International. 14. 63-70 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Keiko Mizuta: "An activated S6 kinase in regenerating rat lever." Biochem. Biophys. Res. Commun.146. 239-246 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Youichirou Sakanoue: "Comparative studies on phosphorylation of synthetic peptide analogue of ribosomal protein S6 and 40S ribosomal subunits between Ca^<2+>-phospholipid-dependent protein kinase and its protease-activated form." European Journal of Biochemistry. 168. 669-677 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Youichirou Sakanoue: "Insulin-stimulated serine kinase in Xenopus oocyto plasma membrane." Biochem. Biophys. Res. Commun.150. 1176-1184 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Kaoru Nishiyama: "Comparison of phosphorylation sites in protamine between protein kinase C and cAMP-dependent protein kinase." Biochemistry International. 17. 51-58 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Eikichi Hashimoto: "Ionic strength-dependent proteolytic activation of protein kinase C by trypsin-like protease." Journal of Biochemistry. 104. 934-938 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Masahiko Hoshijima: "Phosphorylation by cAMP-dependent protein kinase of a human platelet Mr. 22000 GTP-binding protein (Smg p21) having the same putative effector domain as the rat gene products." Biochem. Biophys. Res. Commun.157. 851-860 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Kaoru,Nishiyama: Biochemistry International. 17. 51-58 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Eikichi,Hashimoto: Journal of Biochemistry. 104. 934-938 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Masahiko,Hoshijima: Biochem.Biophys.Res.Commun.157. 851-860 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Akira Takeda: FEBS Letters. 210. 169-172 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Keiko Sakai: Biochemistry International. 14. 63-70 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Keiko Mizuta: Biochem. Biophys. Res. commun.146. 239-246 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Youichiro Sakanoue: European Journal of Biochemistry. 168. 669-677 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Youichiro Sakanoue: Biochem. Biophys. Res. Commun.150. 1176-1184 (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] Eikichi Hashimoto: Biochem. Biophys. Res. Commun.

    • Related Report
      1987 Annual Research Report

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Published: 1987-03-31   Modified: 2016-04-21  

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