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1999 Fiscal Year Final Research Report Summary

Study of the Structure-Function of IPィイD23ィエD2 Receptor/CaィイD12+ィエD1 Release Channel and IPィイD23ィエD2/CaィイD12+ィエD1 Signaling

Research Project

Project/Area Number 10490007
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 広領域
Research InstitutionThe Institute of Physical and Chemical Research (RIKEN) (1999)
The University of Tokyo (1998)

Principal Investigator

FURUICHI Teiichi  The Institute of Physical and Chemical Research (RIKEN), Laboratory Head, 分子神経形成研究チーム, チームリーダー(研究職) (50219094)

Project Period (FY) 1998 – 1999
KeywordsIPィイD23ィエD2 receptor / CaィイD12+ィエD1 signaling / Calmodulin / Ligand binding / Apoptosis / Single channel recording / Cytokinesis / Caspase-3
Research Abstract

1. We found that type 1 IPィイD23ィエD2 receptor-channel, which is abundant in mouse cerebellar microsomal membrane fractions, is composed of five folding domains unsusceptible to limited trypsin digestion. By trypsin digestion, the N-terminal IPィイD23ィエD2-binding region, middle regulatory region and C-terminal channel region were split into 2,3, and one domains, respectively. Trypsin sensitive sites are mainly localized at the alternative splicing sites and regulatory sites such as a calmodulin-binding site, and seem to be exposed to surface of its molecule as a relatively relaxed structure.
2. These five domains generated by trypsin digestion assembled each other as a relatively stable complex through protein-protein interaction, and retained sufficient activities for IPィイD23ィエD2 binding and IPィイD23ィエD2-induced CaィイD12+ィエD1 release. Thus, IPィイD23ィエD2 receptor on CaィイD12+ィエD1 store membranes has a relatively compact folding higher structure, coupling of IPィイD23ィエD2 ligand binding with chann … More el opening may occur through such a tight domain-domain interaction.
3. Of two trypsin-resistant domains containing the IPィイD23ィエD2 binding region, the N-terminal domain had no IPィイD23ィエD2 binding activity, whereas the C-terminal domain exhibited low affinity. On the other hand, by just mixing these two domains expressed individually in E. coli cells we could reconstituted a high-affinity binding activity. These results indicate that these two individual domains can form an active IPィイD23ィエD2 ligand-binding pocked through a protein-protein interaction. We have developed a system to efficiently produce a large amount of soluble protein with high-affinity IPィイD23ィエD2 binding activity in E. coli cells.
4. We analyzed cerebellar type 1 IPィイD23ィエD2 receptor-channel reconstituted into planar lipid bilayers by a single channel recording. As to a biphasic CaィイD12+ィエD1 sensitivity of cerebellar IPィイD23ィエD2 receptor that is thought to deeply be related to intracellular CaィイD12+ィエD1 dynamics, we clarified that an inhibitory phase by about 0.5 μM or higher CaィイD12+ィエD1 concentrations is dependent on CaィイD12+ィエD1-activated calmodulin. These data indicate that calmodulin plays an indispensable role in feedback regulation in intracellular CaィイD12+ィエD1 dynamics.
5. We demonstrated that the local movement of IPィイD23ィエD2 receptor-sensitive intracellular CaィイD12+ィエD1 stores occur during cell division and appears to be involved in CaィイD12+ィエD1 signaling required for cytokinesis.
6. We elucidated that upon induction of T-cell apoptosis the type 1 IPィイD23ィエD2 receptor but not type 2 and type 3 is proteolyzed in dependence on caspase-3, thereby resulting in inactivation of IPィイD23ィエD2 receptor-channel activity. Less

  • Research Products

    (45 results)

All Other

All Publications (45 results)

  • [Publications] Yamamoto-Hino, M.: "Apical vesicles bearing inositol 1, 4, 5-trisphosphate receptor in the Ca^<2+> inhibitation site of ductal epithelium of submandibular gland"J. Cell Biol.. 141. 135-142 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Monkawa, T.: "Localization of inositol 1, 4, 5-trisphosphate receptor in the rat kidney"Kindney Int.. 53. 296-301 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hirota, J.: "T-cell-receptor sinalliing in inositol 1, 4, 5-trisphosphate receptor (IP_3R) type-1-deficient mice : is IP_3R type 1 essential for T-cell-receptor signalling?"Biochem. J.. 333. 615-619 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Zhu, C. C.: "Inositol 1, 4, 5-trisphosphate receptor down- regulation is activated directly by inositol 1, 4, 5-trisphosphate binding"J. Biol. Chem.. 274. 3476-3484 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yoshikawa, F.: "Trysinized cerebllar inositol 1, 4, 5-trisphosphate receptor structural and functional coplin of cleaved ligand binding and channel domains"J. Biol. Chem.. 274. 316-327 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yoshikawa, F.: "Cooperative formation of the ligand binding site of the inositol 1, 4, 5-trisphosphate receptor by two separable domains"J. Biol. Chem.. 274. 328-334 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yoshikawa, F.: "High efficient expression of the functional ligand binding site of the inositol 1, 4, 5-trisphosphate receptor in Escherichia coli"Biochem. Biophys. Res. Commun.. 257. 792-797 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Natsume, T.: "Real time analysis of interaction between inositol 1, 4, 5-trisphosphate receptor type 1 and its ligand"Biochem. Biophys. Res. Commun.. 260. 527-533 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Konishi, Y.: "Transcriptinal regulation of mouse type 1 inositol 1, 4, 5-trisphosphate receptor gene by NeuroD-related factor"J. Neurochem.. 72. 1717-1724 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Mitsuyama, F.: "Microinjection of Ca^<2+> store-enriched microsome fraction to dividing newt eggs induces extra-cleavage furrows via inositol 1, 4, 5-trisphosphate-induces Ca^<2+> release"Dev. Biol.. 214. 160-167 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hirota, J.: "Calmodulin inihibits inositol 1, 4, 5-trisphosphate-induces calcium relase through the purified and reconstituted inositol 1, 4, 5-trisphosphate receptor type 1"FEBS Lett.. 456. 322-326 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Michikawa, T.: "Calmodulin mediates calcium-dependent inactivation of the cerebellar type 1 inositol 1, 4 ,5-trisphosphate receptor"Neuron. 23. 799-808 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shiraishi, Y.: "Cupidin, an isofoem of Homer/Vesl, interacts with the a actin cytoskeleton and activated Rho family small GTPases and is expressed in develping mouse cerebellar granule cells"J. Neurosci.. 18. 8389-8400 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hirota, J.: "Inositol 1, 4, 5-trisphosphate receptor type 1 is a substrate for caspase-3 and is cleaved during apoptosis in a caspase-3 dependent manner"J. Biol. Chem.. 274. 34433-34437 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Baylis, H. A: "Inositol 1, 4, 5-trisphosphate receptor are strongly expressed in the nervous system, pharynx, intestine, gonad and excretory cell of Canorhabditis elegans and are encoded bya single gene (itr-1)"J. Mol. Biol.. 294. 467-476 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Furuichi, T.: "Intracellular Calucium channels."Calcium as a Cellular Regulator. 49 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miyawaki, A.: "Expression of green fluorescent protein and Inositol 1,4,5-triphosphate receptor in xenopus laevis oocytes."Methods in Enzymology. 9 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Mikoshiba, K.: "Role of Inositol 1,4,5-triphosphate(IP_3) receptor /Ca^<2+> signaling in cell function."Challenges for Neuroscience in the 21st Century. 26 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 古市貞一: "イノシトール三リン酸受容体の構造と機能"胃と透析. 8 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 古市貞一: "細胞内Ca遊離チャネルの構造・機能とその病態"Mebio. 7 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 古市貞一: "IP_3受容体と細胞内Ca^<2+>シグナル伝達"日本応用酵素協会誌. 7 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 古市貞一: "カルシウムリリースカルシウムチャンネル"脳の科学. 10 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 竹島浩: "細胞内Ca^<2+>放出チャネル"現代医療. 8 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 御子柴克彦: "IP_3リセプター"蛋白質核酸酵素. 7 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 竹島浩: "カルシウムリリースチャンネルのノックアウト"脳の科学. 6 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 白石陽子: "小脳神経回路の生後発達期に発現変動する遺伝子群"蛋白質核酸酵素. 7 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 古市貞一: "カルシウムチャネル"Clinical Neuroscince(印刷中).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamamoto-Hino: "Apical vesicles bearing inositol 1,4,5-trisphosphate receptors in the CaィイD12+ィエD1 initiation site of ductal epithelium of submandibular gland"J. Cell Biol.. 141. 135-142 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Monkawa,T.: "Localization of inositol 1,4,5-trisphosphate receptors in the rat kidney"Kidney Int.. 53. 296-301 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hirota,J.: "T-cell-receptor signalling in inositol 1,4,5-trisphosphate receptor (IPィイD23ィエD2R) type -1-deficient mice : is IPィイD23ィエD2R type 1 essential for T-cell-receptor signalling?"Biochem. J.. 333. 615-619 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Zhu,C.C.: "Inositol 1,4,5-trisphosphate receptor down-regulation is activated directly by inositol 1,4,5-trisphosphate binding"J. Biol. Chem.. 274. 3476-3484 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoshikawa,F.: "Trypsinized cerebellar inositol 1,4,5-trisphosphate receptor : structural and functional coupling of cleaved ligand binding and channel domains"J. Biol. Chem.. 274. 316-327 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoshikawa,F.: "Cooperative formation of the ligand binding site of the inositol 1,4,5-trisphosphate receptor by two separable domains"J. Biol. Chem.. 274. 328-334 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoshikawa,F.: "High efficient expression of the functional ligand binding site of the inositol 1,4,5-trisphosphate receptor in Escherichia coli"Biochem. Biophys. Res. Commun.. 257. 792-797 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Natsume,T.: "Real time analysis of interaction between inositol 1,4,5-Trisphosphate receptor type 1 and its ligand"Biochem. Biophys. Res. Commun.. 260. 527-533 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Konishi,Y.: "Transcriptional regulation of mouse type 1 inositol 1,4,5-trisphosphate receptor gene by NeuroD-related factor"J. Neurochem.. 72. 1717-1724 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mitsuyama,F.: "Microinjection of CaィイD12+ィエD1 store-enriched microsome fractions to dividing newt eggs induces extra-cleavage furrows via inositol 1,4,5-trisphosphate-induced CaィイD12+ィエD1 release"Dev. Biol.. 214. 160-167 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hirota,J.: "Calmodulin inhibits inositol 1,4,5-trisphosphate-induced calcium release through the purified and reconstituted inositol 1,4,5-trisphosphate receptor type 1"FEBS Lett.. 456. 322-326 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Michikawa,T.: "Calmodulin mediates calcium-dependent inactivation of the cerebellar type 1 inositol 1,4,5-trisphosphate receptor"Neuron. 23. 799-808 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shiraishi,Y.: "Cupidin, an isoform of Homer/Vesl, interacts with the actin cytoskeleton and activated Rho family small GTPases and is expressed in developing mouse cerebellar granule cells"J. Neurosci.. 19. 8389-8400 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hirota,J.: "Inositol 1,4,5-trisphosphate receptor type 1 is a substrate for caspase-3 and is cleaved during apoptosis in a caspase-3 dependent manner"J. Biol. Chem.. 274. 34433-34437 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Baylis,H.A.: "nositol 1,4,5-trisphosphate receptors are strongly expressed in the nervous system, pharynx, intestine, gonad and excretory cell of Caenorhabditis elegans and are encoded by a single gene (itr-1)"J. Mol. Biol. 294. 467-476 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Furuichi,T.: "Intracellular calcium channels"Calcium as a cellular regulator, Oxford Univ. Press. 49 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Miyawaki,A.: "Expression of green fluorescent protein and inositol 1,4,5-triphosphate receptor in xenopus laevis oocytes"Methods in Enzymology, Academic Press. 9 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mikoshiba,K.: "Role of inositol 1,4,5-triphosphate (IPィイD23ィエD2)receptor/ CaィイD12+ィエD1 signaling in cell function"Challenges for Neuroscience in the 21st Century, Japan Scientific Societies Press. 26 (1999)

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

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Published: 2001-10-23  

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