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Involvement and the Molecular Mechanisms of Inositol-1.4.5 Trisphosphate Receptor in the Expression of Neuronal Functions

Research Project

Project/Area Number 02670107
Research Category

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

Allocation TypeSingle-year Grants
Research Field General medical chemistry
Research InstitutionOsaka University

Principal Investigator

NIINOBE Michio  大阪大学, たんぱく質研究所, 助手 (80135748)

Project Period (FY) 1990 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1992: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1991: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1990: ¥1,100,000 (Direct Cost: ¥1,100,000)
KeywordsIP_3 Receptor / IP_3 / IP_4 Binding Protein / IP_4 / アルドラーゼ / IP_4BPI / IP_4結合蛋白質 / IP_4受容体 / IP_3受容体, / Ca^<2+>チャンネル
Research Abstract

1,IP_4結合蛋白質(IP_4BP1)に対するポリクローナル抗体を用いて,マウス脳における本蛋白質の局在性について免疫組織化学的に解析したところ,IP_4BP1は中枢神経系全体に存在することが認められ,とくに,嗅球の外網状層,僧帽細胞層,大脳皮質錐体細胞の細胞体と樹状突起,海馬の放射線維層,小脳プルキンエ細胞の細胞体と樹状突起及び糸球体(顆粒細胞と苔状線維のシナプス形成部位)に高い局在を示した。2,精製したIP_4BP1及びIP_3受容体をそれぞれ人工脂質膜に組み込みIP_3あるいはIP_4による^<45>Caの取り込みが認められるか否か調べたところ,IP_3受容体の場合は強い取り込みが認められたが,IP_4BP1に関しては,今回の実験条件下では何らの取り込みも認められなかった。IP_3受容体の場合は抗体を用いたアフィニティークロマトグラフィーで一段階の操作で精製したものを用いたときにのみ取り込みがみられたところから,IP_4BP1についても変性を極力抑えた条件下での精製法を開発する必要が考えられた。3,精製したIP_4BP1のSDS PAGE像では140Kと65Kの2本の分子種が存在する。これらについて,PVDF膜にトランスファーレ,リジルエンドペプチダーゼ消化を行ない,HPLCで分析したところ,ペプチドの溶出パターンが極めて類似しており,両者が一次構造上高いホモロジーを持つものと推測された。また,140K分子については,それぞれのピークについて現在,アミノ酸配列を決定しつつある。4,IP_4結合蛋白質はマウス小脳には少くとも3種類存在することを既に明らかにしているが,IP_4BP1以外の分子種の一つを完全精製し,N未端からのアミノ酸配列を調べた結果,アルドラーゼのAタイプと同じ配列を持つことが認められた。アルドラーゼはIP_3とも強いアフィニティーを持つことが報告されており,アルドラーゼの細胞内機能にIP_3及びIP_4がどのように関わりを持つのか現在,検討しつつある。

Report

(4 results)
  • 1992 Annual Research Report
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • Research Products

    (21 results)

All Other

All Publications (21 results)

  • [Publications] Maeda,N.: "A Cerebellar Purkinje cell marker P400 protein is an inesitol 1,4,5-trisphosphate(Ins P_3)receptor protein.Purification and characterization of Ins P_3 receptor conplex." EMBO J.9. 61-67 (1990)

    • Related Report
      1992 Annual Research Report
  • [Publications] Mikoshiba,K.: "Primary structure and functional expression of the inositol 1,4,5-trisphosphate reoptor,P400." Neuroreceptor Mechanisms in Brain.Edited by S.Kito et al.,Plenum press,New York. 83-95 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Kuwashima,G.: "Two types of rianodine receptors in mouse biain:Skeletal muscle type exclusinely in Purkinje cells and cardioc muscle type in ranous neurons." Neuron. 9. 1133-1142 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Maeda,N: "A cerebellar Purkinje cell marker P400 protein is an inositol 1,4,5-trisphosphate(InsP_3)receptor protein.Purification and characterization of InsP_3 receptor complex." EMBO J.9. 61-67 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Mikoshiba,K.: "Primary structure and functional expression of the inositol 1,4,5-trisphosphate receptor,P400." Neuroreceptor Mechanisms in Brain,Edited by S. Kito et al.,plenum press,New York. 83-95 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kuwashima,G.: "Two types of rianodine receptors in mouse brain:skeletal muscle type exclusively in Purkinje cells and cardiac muscle type in various neurons." Neuron. 9. 1133-1142 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Maeda, N: "A cerebellar Purkinje cell marker P400 protein is an inositol 1, 4, 5-trisphosphate (InsP_3) receptor protein. Purification and characterization of InsP_3 receptor complex" EMBO J. 9(1). 61-67 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Mikoshiba, K: "Primary structure and functional expression of the inositol 1, 4, 5-trisphosphate receptor, P400" Neuroreceptor Mechanisms in Brain, Edited by S. Kito et al., Plenum press, New York. 83-95 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kuwashima, G: "Two types of rianodine receptors in mouse brain: skeletal muscle type exclusively in Purkinje cells and cardiac muscle type in various neurons" Neuron. 9. 1133-1142 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Yamamoto,H.: "Phosphorylation of P400 protein by cyclic AMPーdependent protein kinase and Ca^<2+>/calmodulinーdependent protein kinase 2." J.Neurochem.53. 917-923 (1989)

    • Related Report
      1991 Annual Research Report
  • [Publications] Maeda,N.: "Oevelopmental expression and intracellulan locntion of P400 protein characteristic of Puskinje cells in the mouse cerebellum." Develop.Biol.133. 67-76 (1989)

    • Related Report
      1991 Annual Research Report
  • [Publications] Maeda,N.: "A cevebellan Puskinze cell marker P400 protein is an inositol 1,4,5ーtn3phosphate(Ins P_3)receptor protein,Pusfication and characterization of Ins P_3 receptor complex." EMBO J.9. 61-67 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Otsu,H.: "Immunogold localization of Inositol 1,4,5ーtrisphosplate(IP_3)receptor in mouse cerebellen Puskinge using three monoclonal antibodies." Cell Structure and Function. 15. 163-173 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Maeda,N.: "Structural and functional cheractesization of Inositol 1,4,5ーtrisphosphate receptor chnnel from mouse cerebellum." J.Biol.Chem.266. 1109-1116 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Nakade,S.: "Involvement of the Cーterminas of the inositol 1,4,5ーtrisphophate receptor in Ca^<2+>release analysed using regionーspecific monoclonal antibodies." Biochem.J.277. 125-131 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Maeda,N.: "Purification and characterization of P400 profein,a glycoprotein characteristic of Purkinje cell from mouse cerebellum." J.Neurochem.51. 1724-1730 (1988)

    • Related Report
      1990 Annual Research Report
  • [Publications] Yamamoto,H.: "Phosphorylation of P400 proteim by cyclic AMPーdependent protein kinase and Ca^<2+>/calmodulinーdependent proteim kinaseII." J.Neurochem.53. 917-923 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] Maeda,N.: "Developmental expression and intracellulan location of P400 protein characteristic of Purkinje cells in the mouse cerebellum." Develop. Biol.133. 67-76 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] Maeda,N.: "A cerebellan Purkinje cell marker P400 Protein is an inositol 1,4,5ーtrisphosphate(InsP_3)receptor protein,Purification and characterization of InsP_3 receptor complex." EMBO J.9. 61-67 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Otsu,H: "Immurogold localization of Inositol 1,4,5ーtrisphosーphote(InsP_3)receptor in mouse orebellar Purkinje cells using three monocloral antiboclies." Cell Structure and Function. 15. 163-173 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Maeda,N: "Structural and functional characterization of Inositol 1,4,5ーtrisphosphate receptor channel from mouse cerebellum." J.Biol. Chem.266. 1109-1116 (1991)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2017-05-19  

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