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Structure and Function of Protein Kinases: Molecular Mechanism of Neuronal Signal Transduction

Research Project

Project/Area Number 63440021
Research Category

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

Allocation TypeSingle-year Grants
Research Field General pharmacology
Research InstitutionKobe University School of Medicine

Principal Investigator

TANAKA Chikako  Kobe Univ. Sch. Med., Dept. of Pharmacol., Professor, 医学部, 教授 (20025571)

Co-Investigator(Kenkyū-buntansha) KUNO Takayoshi  Kobe Univ. Sch. Med., Dept. of Pharmacol., Assistant Professor, 医学部, 講師 (50144564)
TANIYAMA Kohtaro  Kobe Univ. Sch. Med., Dept. of Pharmacol., Associate Professor, 医学部, 助教授 (70030898)
Project Period (FY) 1988 – 1989
Project Status Completed (Fiscal Year 1989)
Budget Amount *help
¥12,100,000 (Direct Cost: ¥12,100,000)
Fiscal Year 1989: ¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 1988: ¥8,000,000 (Direct Cost: ¥8,000,000)
KeywordsProtein kinase C / cAMP-dependent protein kinase / Synaptic transmission / Neurotransmission / Cellular signal transduction / Immunohistochemistry / プロテインキナーゼC / サブタイプ / cAMP依存性プロテインキナーゼ / 制御サブユニット / cAMP結合部位
Research Abstract

The importance of protein kinases as target molecules of second messengers in the cellular signal transduction mechanisms activated by hormones and neurotransmitters are well established. Protein kinase C and cAMP-dependent protein kinase are thought to be major protein kinases in these mechanisms. In this study, we tried to elucidate the molecular mechanisms of neuronal signal transduction related to these protein kinases.
1) Localization and function of protein kinase C isozymes: Light- and electron-microscopic localization of alpha, beta-I, beta-II, and gamma subtypes of protein kinase C in rat brain were elucidated using specific antibodies. Immunohistochemical analysis of developing rat brain showed that each subtype expresses at different stage. Isozymes related to neurorite outgrowth and synaptic formation were identified. Specific expression of these isozymes was also shown by the in situ hybridization technique.
2) The role of protein kinase C in the synaptic transmission: Invol … More vement of protein kinase C in calcium-dependent vesicular neurotransmitter release was shown. Gamma subtype of protein kinase C which is activated by arachidonic acid was shown to be involved in the GABA release from Purkinje cell in cerebellum.
3) Molecular cloning and expression of regulatory subunit of cAMP-dependent protein kinase: A cDNA clone containing an entire coding region of the regulatory subunit was cloned from a rat brain cDNA library. Wild or mutant regulatory subunits were expressed in E. coli. After incubation with catalytic subunit purified from bovine heart, holo-enzymes were activated by CANW. Although these subunits bound cAMP with high affinity, holo-enzymes contains mutant subunits could not be activated by high concentrations of cAMP, indicating the importance of the presence of intrachain two cAMP-binding sites. These studies suggest that subtypes of protein kinase C have specific roles in neurorite outgrowth, synaptic formation, neurotransmitter release and long term potentiation, and showed the importance of protein phosphorylation in the neuronal signal transduction. Less

Report

(3 results)
  • 1989 Annual Research Report   Final Research Report Summary
  • 1988 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] A.Ito: "Immunocytochemical Iocalization of α-subspecies of protein kinase C in rat brain." Proc.Natl.Acad.Sci.USA.(in press). (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] K.Hosoda: "Immunocytochemical Iocalization of the βI-subspecies of protein kinase C in rat brain." Proc.Natl.Acad.Sci.USA..86. 1393-1397 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] N.Saito: "Immunocytochemical Iocalization of βII-subspecies of protein kinase C in rat brain." Proc.Natl.Acad.Sci.USA.86. 3409-3431 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] A.Ito: "Localization of βII-subspecies of protein kinase C in B cell of pancreatic islet." Diabetes. 38. 1005-1011 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] T.Tsujino: "Light and electron microscopic localization of βI-,βII-and γ-subspecies of protein kinase C in rat cerebral cortex." J.Neurosci.(in press). (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] A.Kose: "Electron microscopic localization of γ-and βII-subspecies of protein kinase C in rat hippocampus." Brain Res.(in press). (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] A. Ito: "Immunocytochemical localization of alpha-subspecies of protein kinase C in rat brain." Proc. Natl. Acad. Sci. USA.(1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] K. Hosoda: "Immunocytochemical localization of the betaI-subspecies of protein kinase C in rat brain." Proc. Natl. Acad. Sci. USA. 86. 1393-1397 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] N. Saito: "Immunocytochemical localization of betaII-subspecies of protein kinase C in rat brain." Proc. Natl. Acad. Sci. USA. 86, 1989, 3409-3431.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] A. Ito: "Localization of betaII-subspecies of protein kinase C in B cell of pancreatic islet." Diabetes 38, 1989, 1005-1011.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] T. Tsujino: "Light and electron microscopic localization of betaI-, betaII- and gamma-subspecies of protein kinase C in rat cerebral cortex." J. Neurosci. 1990.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] A. Kose: "Electron microscopic localization of gamma- and betaII- subspecies of protein kinase C in rat hippocampus." Brain Res. 1990.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] A.Ito: "Immunocytochemical localization of alpha-subspecies of protein kinase C in rat brain." Proc.Natl.Acad.Sci.USA.in press. (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] K.Hosoda: "Immunocytochemical localization of the betaI-subspecies of protein kinase C in rat brain." Proc.Natl.Acad.Sci.USA. 86. 1393-1397 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] N.Saito: "Immunocytochemical localization of betaII-subspecies of protein kinase C in rat brain." Proc.Natl.Acad.Sci.USA.86. 3409-3431 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] A.Ito: "Localization of betaII-subspecies of protein kinase C in B cell of pancreatic islet." Diabetes. 38. 1005-1011 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] T.Tsujino: "Light and electron microscopic localization of betaI-,betaII- and gamma- subspecies of protein kinase C in rat cerebral cortex." J.Neurosci.(1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] A.Kose: "Electron microscopic localization of gamma- and betaII- subspecies of protein kinase C in rat hippocampus." Brain Res.(1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] N.Saito: Pro.Natl Acad.Sci.USA. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] K.Hosoda: Pro.Natl.Acad.Sci.USA. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] A,Kose: J.Neurosci.8. 4262-4268 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] H.Shuntoh: J.Neurochem.51. 1565-1572 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] T.Kuno: Biochem.Biophys.Res.Commun.153(3). 1244-1250 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] N.Saito: 8. 369-382 (1988)

    • Related Report
      1988 Annual Research Report

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Published: 1988-04-01   Modified: 2016-04-21  

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