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2000 Fiscal Year Final Research Report Summary

Studies on the functions of a novel inositol 1, 4, 5-trisphosphate binding protein

Research Project

Project/Area Number 11694288
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field General medical chemistry
Research InstitutionKYUSHU UNIVERSITY

Principal Investigator

HIRATA Masato  KYUSHU UNIVERSITY Faculty of Dental Science Prof., 歯学研究院, 教授 (60136471)

Co-Investigator(Kenkyū-buntansha) MATILDA Katan  連合王国癌研究所, チェスタービーティーラボラトリー, 主任研究員
TAKEUCHI Hiroshi  KYUSHU UNIVERSITY Faculty of Dental Science Assistant Prof., 歯学研究院, 助手 (70304813)
KANEMATSU Takashi  KYUSHU UNIVERSITY Faculty of Dental Science Associate Prof., 歯学研究院, 助教授 (10264053)
KATAN Matilda  Chester-Beatty Lab, (UK) Principle Investigator
Project Period (FY) 1999 – 2000
Keywordscalcium ion / inositol 1, 4, 5-trisphospahte / gamma-aminobutyric acid / central nervous system / anti-anxiety drug / protein phosphatase
Research Abstract

We isolated a novel inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) binding proteins with a molecular mass of 130kDa (p130 or PRIP1), which was later found to be a similar protein to delta-isozyme of phospholipase C, but with no catalytic activity. Recently, we established a cell-line, stably over-expressing p130 (COS-1p130) in order to assess the physiological function. Fura-2-loaded COS-1p130 were stimulated with either bradykinin (BK) or epidermal growth factor (EGF) and the changes in free Ca2+ concentration was monitored. Compared to the control cells, the responses of free Ca2+ change were diminished without the changes of levels of Ins(1,4,5)P3 production in response to BK and EGF, indicating that the diminution of Ca2+ response by p130 could be a result of binding of cellular Ins(1,4,5)P3 produced by PLC activation. The p130 might be a negative regulator for Ca2+ signaling pathways in cells. We further attempted to identify interacting molecules to help elucidate the function of p130. We isolated two clones interacting with p130 by yeast two-hybrid screening, the sequencing of which revealed that one was protein phosphatase 1 (PP1) catalytic subunit and the other was GABARAP (GABAA receptor associated protein). We then investigated the region responsible for their interactions and found that PP-1 and GABARAP associated with the pleckstrin homology domain (PH domain) and EF-hand motifs of p130, respectively. To elucidate the function of p130 on GABAA receptor signaling, the effect of zinc ion on IGABA was investigated by whole cell patch recording using acutely dissociated hippocampal CA1 neurons from p130-/- mice. The inhibitory action of zinc ion on IGABA decreased significantly in p130-/-mice. These results suggest that p130 is involved in signaling pathway via not only an Ins(1,4,5)P3-Ca2+ system but also a GABAA receptor signaling concerning with PP1 and GABARAP.

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Takeuchi,H. et al: "Membrane association of a new inositol 1,4,5-trisphosphate binding protein, p130 is not dependent on the pleckstrin homology domain"Chem.Phys.Lipids. 98. 35-47 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Matsuki,N. et al: "Antibodies against the PH domain of phospholipase C-d1 inhibit Ins (1,4,5)P3-mediated Ca2+ release from the endoplasmic reticulum"Biochem.Biophys.Res.Commun.. 260. 42-47 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamamoto,T. et al: "Involvement of EF hand motifs in the Ca2+-dependent binding of the pleckstrin homology domain to phosphoinositides"Eur.J.Biochem.. 265. 481-490 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kanematsu,T. et al: "Domain organization of p130, PLC-related catalytically inactive protein, and structural basis for the lack of enzyme activity"Eur.J.Biochem.. 267. 2731-2737 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takeuchi,H. et al: "Inhibition of calcium signalling by p130, PLC-related catalytically inactive protein : critical role of the p130 PH domain"Biochem.J.. 349. 357-368 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hermosura,M.C. et al: "InsP4 facilitates store-operated calcium influx by inhibition of InsP35-phosphatase"Nature. 408. 735-740 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takeuchi, H., Kanematsu, T., Misumi, Y.and Hirata, M.: "Membrane association of a new inositol 1, 4, 5-trisphosphate binding protein, p130 is not dependent on the pleckstrin homology domain."Chem.Phys.Lipids.. 98. 35-47 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Matsuki, N., Tateishi, K., Takeuchi, H., Yagisawa, H., Kanematsu, T., Oishi, M, and Hirata, M.: "Antibodies against the PH domain of phospholipase C-d1 inhibit Ins(1, 4, 5)P3-mediated Ca2+release from the endoplasmic reticulum."Biochem.Biophys.Res.Commun.. 260. 42-47 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamamoto, T., Takeuchi, H., Kanematsu, T., Allen, V., Yagisawa, H., Kikkawa, U., Watanabe, Y., Nakasima, A., Katan, M.and Hirata, M.: "Involvement of EF hand motifs in the Ca2+-dependent binding of the pleckstrin homology domain to phosphoinositides."Eur.J.Biochem.. 265. 481-490 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kanematsu, T., Yoshimura, K., Hidaka, K., Takeuchi, H., Katan, M.and Hirata, M.: "Domain organization of p130, PLC-related catalytically inactive protein, and structural basis for the lack of enzyme activity."Eur.J.Biochem.. 267. 2731-2737 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takeuchi, H., Oike, M., Paterson, H.F., Allen.V., Kanematsu, T., Ito, Y., Erneux, C., Katan, M.and Hirata, M.: "Inhibition of calcium signalling by p130, PLC-related catalytically inactive protein : critical role of the p130PH domain."Biochem.J.. 349. 357-368 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hermosura, M.C., Takeuchi, H., Fleig, A., Riley, A.M., Potter, B.V.L., Hirata, M.and Penner, R.: "InsP4 facilitates store-operated calcium influx by inhibition of InsP3 5-phosphatase"Nature. 408. 735-740 (2000)

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

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Published: 2002-03-26  

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