• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Regulation of neuronal cell ion channels by G proteins

Research Project

Project/Area Number 06044079
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research InstitutionTOKYO INSTITUTE OF TECHNOLOGY

Principal Investigator

ITOH Hiroshi  TOKYO INSTITUTE OF TECHNOLOGY, 生命理工学部, 客員助教授 (10183005)

Co-Investigator(Kenkyū-buntansha) MIZUNO Norikazu  TOKYO INSTITUTE OF TECHNOLOGY, 生命理工学部, 寄附講座教員 (90212232)
KOZASA Tohru  UNIVERSITY OF TEXAS WE MEDICAL CTR, 医学部, 助教授
NAKAJIMA Yasuko  UNIVERSITY OF ILLINOIS AT CHICAGO, 医学部, 教授
KAZIRO Yoshito  TOKYO INSTITUTE OF TECHNOLOGY
NAKAJIMA Shigehiro  UNIVERSITY OF ILLINOIS AT CHICAGO
KAWANO Takeharu  TOKYO INSTITUTE OF TECHNOLOGY
SATOH Takaya  TOKYO INSTITUTE OF TECHNOLOGY
佐藤 孝哉  東京工業大学, 生命理工学部, 寄附講座教員 (20251655)
上代 淑人  東京工業大学, 生命理工学部, 客員教授 (90012690)
Project Period (FY) 1994 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥10,500,000 (Direct Cost: ¥10,500,000)
Fiscal Year 1996: ¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1995: ¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1994: ¥3,700,000 (Direct Cost: ¥3,700,000)
KeywordsG PROTEINS / ION CHANNELS / NEURON
Research Abstract

Heterotrimeric GTP-binding regulatory proteins (G proteins) mediate a variety of extracellular signals from receptors to effector molecules. G protein-coupled inward rectifier K+ channels (GIRKs) are known to be one of the effector molecules, and are involved in the inhibition of neuronal excitability and the development of central neurvous system. To investigate the regulation mechanism of GIRK by G proteins, we examined the heteromultimeric complex formation of different GIRK and the interaction of the GIRKs with G protein betagamma subunits using co-immunoprecipitation technique. We cloned cDNAs coding for three types of GIRKs, GIRK1, GIRK2, and GIRK4, from rat brain. The GIRK cDNAs were subcloned into mammalian expression vector, and expressed in human embryonal kidney 293 cells. Using specific antibody against each GIRK,the GIRK was immunoprecipitated. The immunoprecipitate was analyzed by immunoblotting. The results of co-immunoprecipitation of the two different GIRKs in three different combination indicated that GIRK1, GIRK2, and GIRK4 intreract each other. to from a heteromultimer complex. Interaction of GIRK with G protein betagamma subunit (Gbetagamma) was studied using various mutants of Gbeta and Ggamma. We found that the association of GIRK with Gbetagamma is mediated mainly through Gbeta, but not Ggamma, and Gbeta alone can interact with GIRK.Moreover, it was suggested that the membrane localization of Gbetagamma is required for association of GIRK.Futhermore, we succeeded in measuring the activity of GIRK which was transiently expressed together with Gbetagamma in 293 cells and in primary cultured neuron. Pilot experiments using the Gbeta mutants suggested that the association of GIRK with Gbeta alone fails to activate the GIRK and the Gbetagamma dimer functions as an activator of GIRK.Futher studies including electrophysical and molecular biological experiments should throw a new linght on the molecular mechanisms of regulation of GIRK.

Report

(4 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • 1994 Annual Research Report
  • Research Products

    (44 results)

All Other

All Publications (44 results)

  • [Publications] H.Edamatsu et al.: "Inducible high-level expression rector for mammalian cells,pEF-LAC carrying human elongation factor 1α promoter and lac operator" Gene. (in press). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] J.Yamauchi et al.: "C-terminal mutation of G protein β subunit affects differentially extracellular signal-regulated kinase andc-Jun N-terminal kinase pathways in HEK 293 cells" Journal of Biological Chemistry. (in press). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Y.Nakajima et.al: "Activation of G protein-coupled inward rectifier K^+ channels in brain neurons requires association of G protein βγ subunits with cell membrane" FEBS Letters. 390. 217-220 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] R.Farkas et al: "Properties of a slow nonselective cation conretence modulated by neurotensin and other neurotransmitters in midbrain dopaminergic neurons" Journal of Neurophysiology. 76.3. 1968-1981 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] P.Chien et al.: "Single channel propeties of the nonselective cation condretance induced by neurotensin in dopaminergic neurons" Proceedings of the National Academy of Sciences of the United States of America. 93.25. 14917-14921 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] K.Takano et al: "G q/11 and PLC-β1 mediate the substance P-induced inhibition of an inward rectifier K^+ channel in brain neuron" Journal of Neurophysiology. 76.3. 2131-2136 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 伊東 広 他: "GTP結合蛋白質" 羊土社, 312 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 伊東 広 他: "脳における情報伝達" 共立出版, 608 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 伊東 広 他: "理化学辞典" 岩波書店, 1650 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] J.Yamauchi, Y.Kaziro, and H.Itoh: "C-terminal mutation of G protein beta subunit affects differentially extracellular signal-regulated kinase and c-Jun N-terminal kinase pathways in HEK 293 cells" Jounal of Biological Chemistry. 272(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Edamatsu, Y.kaziro, and H.Itoh: "Inducible high-level expression vector for mammalian cells, pEF-LAC carrying human elongation factor 1alpha promoter and lac operator" Gene. 180(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] K.Horie, H.Itoh, and G.Tsujimoto: "Hamster alpha1B-adrenergic receptor directly activates Gs in the transfected CHO cells" Molecular Pharmacology. 48. 392-400 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] A.Ito, T.Satoh, Y.Kaziro, and H.Itoh: "G protein betagamma subunit activates Ras, Raf, and MAP kinase in HEK 293 cells." FEBS Letters. 365. 183-187 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] P.Chien, R.Farkas, S.Nakajima, and Y.Nakajima: "Single channel properties of the nonselective cation conductance induced by neurotensin in doparminegic neurons" Proc.Natl.Acad.sci.U.S.A.93. 14917-14921 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] K.Takano, P.R.Stanfield, S.Nakajima, and Y.Nakajima: "Protein kinase C-mediated inhibition of an inward rectifier potassium channel by substance P in nucleus basalis neurons." Neuron. 14. 999-1008 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] B.M.Velimirovic, K.Koyano, S.Nakajima, and Y.Nakajima: "Opposing mechanisms of regulation of a G-protein-coupled inward rectifier K+ channel in rat brain neurons" Proc.Natl.Acad.Sci.U.S.A.92. 1590-1594 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Itoh: Kyoritsu-shuppan. Structure and regulation of adenylyl cyclase, 371-378 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Itoh, and J.Yamauchi: Yodosya. Gene and structure of G protein subunits, 30-37 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Itoh: Hirokawa publishing Co.Superfamily of G protein genes, 93-103 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Itoh: Ishiyaku publishing Co.Signal transduction by G proteins, 337-342 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Y.Nakajima, and S.Nakajima: Humana Press Inc.signal transduction mechanisms of tachykinin effects on ion channels., 285-327 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Edamatsu et al.: "Inducible High-lerel expression vector for mammalian cells,PEF-LAC carrying human elougation factor 1d promoter and lac operator" Gene. (in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] J.Yamauchi et al.: "C-terminal mutation of G protein β subunit affects differentially extracellular signal-regulated kinase and c-Jun N-terminal Kinasw pathways in HEX 293cells" Journal of Biological Chemistry22GD02:(in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y.Nakajima et al.: "Activation of G protein-compled inward rectifier k^+ channals in brain neurons regrsives sssoctation of G protein Pr subnits w.th cell membrane" FEBS Letters. 390. 217-220 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] R.Farkas et al.: "Properties of a slow nonselective cation conductance modulated by neurotensin and other neurotensins there in midbrain dopaminegic nerons" Journal of Neurophysiology. 76・3. 1968-1981 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] P.Chien et al.: "Single channel properties of the nonselective cation cnanetance induced by neurotensin in dopaminergic neurons" Proceedings of the National Academy of Sciences of the Uniteol States of America. 93・25. 14917-14921 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.Takano et al.: "Gg/11 and PLC-β1 nediate the substance P-induced inhibition of an inward rectifier k^+ channel in brain neuron" Journal of Neurophysiology. 76・3. 2131-2136 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 伊東広 他: "GTP結合蛋白質" 羊士社, 312 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 伊東広 他: "脳における情報伝達" 共立出版, 608 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 伊東広 他: "理化学辞典" 岩波書店, 1650 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] A.Ito: "G pntein βγ subunit activates Ras,Rsf,and MAP Kinase in HEK 293 cells" FEBS Lett.368. 183-187 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] J.Yamauchi: "Carboxyl terminal of G protein β subunits ireguires for association with γ subunit" Biochem.Biophys.Res.Commun.214. 694-700 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] K.Horie: "Hamster α_<1B>-Adrenergic receptor directly activates G_S in the transfected CHO cells" Molecular Pharmacology. 48. 392-400 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] B.Velimirovic: "Opposing mechanisms of regulation of a G protein-compled inward rectifier K^T channel in rat brain neurons" Proc.Natl.Acad.Sci.USA. 92. 1590-1594 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] K.Takano: "Protein Kinase C-mediated inhibition of an inward rectifier potassium channel by substance P in nucleus basalis neurons" Neuron. 14. 999-1008 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] K.Takano: "Gq/11 mediates substance P-induced inhibition of the inwardly rectifyging K^T current in nucleus basalis neurons" Biophys.J.68. A12- (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 伊東 広: "情報変換物質:Gタンパク質 広川ニューロサイエンス" 広川書店, 166 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 伊東 広: "GTP結合蛋白質" 羊士社, 312 (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] 伊東 広: "G蛋白質を介するシグナル伝達" 医学のあゆみ. 171. 337-342 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ronald H.Farkas et al.: "Neurotensin excites basal forebrain cholinergic newrons: Ionic and signaltransduetion mechanisms" Proc.Natl.Acad.Scin USA. 91. 2855-2857 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ronald H.Forkas et al.: "Cultured neurons infected with an HSV-1-derived vector remain electrically excstable and responsive to neurotransmitter" Neuroscience Letters. 165. 153-156 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 伊藤明子ら.: "Gタンパク質βγサブユニット複合体を介するMAPキナーゼの活性化" 生化学. 66. 751 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 伊東 広: "ニューロサイエンス講座5 情報交換物質:G蛋白" 広川書店, 200 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Yasuko Nakajima: "The Tachykinin Receptors" Humana Press.Inc., 327 (1994)

    • Related Report
      1994 Annual Research Report

URL: 

Published: 1994-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi