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1992 Fiscal Year Final Research Report Summary

Studies on Protein Kinases in Rat Cerebral Postsynaptic Density

Research Project

Project/Area Number 03680160
Research Category

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

Allocation TypeSingle-year Grants
Research Field 代謝生物化学
Research InstitutionAsahikawa Medical College

Principal Investigator

KAMESHITA Isamu  Asahikawa Medical College, Associate Professor, 医学部, 助教授 (60127941)

Co-Investigator(Kenkyū-buntansha) FUJISAWA Hitoshi  Asahikawa Medical College, Professor, 医学部, 教授 (10027039)
Project Period (FY) 1991 – 1992
KeywordsProtein kinase / Calmodulin / Protein phosphorylation / Central nervous system / Signal transduction
Research Abstract

(1) Protein Kinases in Postsynaptic Density---Ten protein kinases of Mr=116kDa, 95kDa, 78kDa, 70kDa, 64kDa, 60kDa, 50kDa, 41kDa, 37kDa, and 32kDa have been detected in the postsynaptic density (PSD) from rat brain by means of the activity gel method. Two protein kinases of Mr=50kDa and 60kDa were identified as the alpha and beta subunits of CaM-dependent protein kinase II (CaM-kinase II), respectively. In addition to CaM-kinase II, CaM-independent protein kinases of Mr=95kDa, 78kDa, 70kDa, and 64kDa appeared to be highly enriched in cerebral PSD fraction. These protein kinases were found to be neutral proteins with pI values ranging from 6.7-7.2 when determined by the two-dimentional gel electrophoresis. However, their natural substrates and physiological functions have not been clarified yet.
(2) Purification and Characterization of Brain-specific CaM- dependent Protein Kinase IV (CaM-kinase IV)---CaM-kinase IV has been purified from rat cerebellum. The enzyme was a monomeric enzyme with a molecular weight of 67,000. The enzyme phosphorylated various substrates such as synapsin I, MAP2, myosin light chain, and tyrosine hydroxylase, suggesting that the enzyme is a multifunctional protein kinase. The enzyme underwent autophosphorylation in a Ca^<2+>/CaM-dependent manner. Approximately one mol of phosphate was incorporated into one mol of the enzyme, resulting in a marked stimulation of the enzyme activity. In contrast, CaM-kinase IV was phosphorylated by cAMP- dependent protein kinase, leading to a significant decrease in the enzyme activity. Thus, CaM-kinase IV appears to be under the control of two important intracellular signalling system, the cAMP system and the Ca^<2+> system.

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] 亀下 勇: "ラット大脳CaM依存性プロテインキナーゼIVの自己リン酸化反応" 神経科学. 30. 212-213 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Isamu KAMESHITA: "Phorphouylation and Functional Modification of CaM-dependeit ProteinkinaseIV by cAMP-dependent Protein Kimase" Biochem.Biophys.Red.Commun. 180. 191-196 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 亀下 勇: "ラット大脳CaM-キナーゼIVのA-キナーゼによるリン酸化と活性調節" 神経科学. 31. 40-41 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Osamu MIYANO: "Pueification and Characterigation of roin-Specific Multifumctional CaM-dependent Protein Kinase from Rat Cerebellun" J.Biol.Chem. 267. 1198-1203 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 亀下 勇: "カルシウム依然性プロティンキナーゼ" Clinical Calcium. 3. 293-298 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Isamu KAMESHITA: "Autophesphorylation of Calmobulim dependent Protein Kinase from Rat Cerebral Cortox" J.Biochem. (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Isamu Kameshita: "Autophosphorylation of CaM-dependent Protein Kinase IV from Rat Cerebral Cortex" Shinkei Kagaku. 30. 212-213 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Isamu Kameshita: "Phosphorylation and Functional Modification of CaM-dependent Protein Kinase IV by cAMP-dependent Protein Kinase" Biochem. Biophys. Res. Commun.180. 191-196 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Isamu Kameshita: "Regulation of CaM-dependent Protein Kinase IV Activity by Phosphorylation with cAMP-dependent Protein Kinase" Shinkei Kagaku. 31. 40-41 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Osamu Miyano: "Purification and Characterization of a Brain-specific Multifunctional CaM-dependent Protein Kinase from Rat Cerebellum" J. Biol. Chem.267. 1198-1203 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Isamu Kameshita: "Calcium-dependent Protein Kinase" Clinical Calcium. 3. 293-298 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Isamu Kameshita: "Autophosphorylation of Calmodulin-dependent Protein Kinase IV from Rat Cerebral Cortex" J. Biochem.(1993)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1994-03-24  

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