1999 Fiscal Year Annual Research Report

IP_3受容体/Ca^<2+>放出チャネルの構造機能とIP_3/Ca^<2+>シグナル伝達の研究

Research Project

Project/Area Number	10490007
Research Institution	The Institute of Physical and Chemical Research (RIKEN)
Principal Investigator	古市貞一理化学研究所, 分子神経形成研究チーム, チームリーダー(研究職) (50219094)
Keywords	IP_3受容体 / Ca^<2+>シグナル伝達 / カルモジュリン / リガンド結合 / アポトーシス / 単一チャネル記録 / 細胞質分裂 / カスパーゼ3
Research Abstract	小脳のタイプ1IP_3受容体のチャネル活性を平面脂質二重膜に再構成して単一チャネル記録解析を行った。その結果、IP_3受容体の二相性Ca^<2+>感受性のうち、約0.5μM以上の高Ca^<2+>濃度によってもたらされる抑制相はカルモジュリンに依存していることを明らかにした。我々は先に、IP_3受容体にカルモジュリン結合部位を見つけているが、今回の結果はカルモジュリンがIP_3受容体の高Ca^<2+>濃度センサーとしてはたらいてIP_3誘導Ca^<2+>放出を抑制することで、細胞内Ca^<2+>ダイナミクスの制御に重要な役割を演じることを示唆している。 IP_3受容体のリガンド結合部位のタンパク質構造をトリプシン感受性などを指標に考察することにより、高親和性のIP_3結合活性を高収率で可溶性の活性タンパク質として大腸菌で発現させるシステムを開発した。細胞分裂に伴ってIP_3受容体感受性の細胞内Ca^<2+>ストアの局在はダイナミックな変動を示し、細胞質分裂に必要なCa^<2+>シグナル伝達に関与することを明らかにした。 T細胞のアポトーシス誘導によってタイプ1IP_3受容体がカスパーゼ3依存的な部分分解を受け、その結果、チャネル活性が不活化されることを明らかにした。但し、タイプ2と3では分解はみられなかった。

Research Products

(19 results)

All Other

All Publications (19 results)

[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)
[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)
[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)
[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)
[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)
[Publications] Baylis H.A.: "Inositol 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"J.Mol.Biol.. 294. 467-476 (1999)
[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)
[Publications] Natsume T.: "Real time analysis of interaction between inositol 1,4,5-Trisphosphate receptor type 1 and its ligand"Biochem.Biophts.Res.Commun.. 260. 527-533 (1999)
[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)
[Publications] Mitsuyama F.: "Microinjection of Ca^<2+> store-enriched microsome fractions to dividing newt eggs induces extra-cleavage furrows via inositol 1,4,5-trisphosphate-induced Ca^<2+> release"Dev.Biol.. 214. 160-167 (1999)
[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)
[Publications] Michikawa T.: "Calmodulin mediates calcium-dependent inactivation of the cerebellar type 1 inositol 1,4,5-trisphosphate receptor"Neuron. 23. 799-808 (1999)
[Publications] Furuichi T.: "Intracellular Calucium channels."Calcium as a Cellular Regulator. 49 (1999)
[Publications] Miyawaki A.: "Expression of green fluorescent protein and inositol 1,4,5-triphosphate receptor in xenopus laevis oocytes"Methods in Enzymology. 9 (1999)
[Publications] Mikoshiba K.: "Role of inositol 1,4,5-trisphosphate (IP_3) receptor/Ca^<2+> signaling in cell function"Challenges for Neuroscience in the 21st Century. 26 (1999)
[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)
[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)
[Publications] 竹島浩: "細胞内Ca^<2+>放出チャネル"現代医療. 8 (1999)
[Publications] 古市貞一: "カルシウムチャネル"Clinical Neuroscience(印刷中).

1999 Fiscal Year Annual Research Report

IP_3受容体/Ca^<2+>放出チャネルの構造機能とIP_3/Ca^<2+>シグナル伝達の研究

Principal Investigator

古市 貞一 理化学研究所, 分子神経形成研究チーム, チームリーダー(研究職) (50219094)

Research Products

[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)

[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)

[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)

[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)

[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)

[Publications] Baylis H.A.: "Inositol 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"J.Mol.Biol.. 294. 467-476 (1999)

[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)

[Publications] Natsume T.: "Real time analysis of interaction between inositol 1,4,5-Trisphosphate receptor type 1 and its ligand"Biochem.Biophts.Res.Commun.. 260. 527-533 (1999)

[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)

[Publications] Mitsuyama F.: "Microinjection of Ca^<2+> store-enriched microsome fractions to dividing newt eggs induces extra-cleavage furrows via inositol 1,4,5-trisphosphate-induced Ca^<2+> release"Dev.Biol.. 214. 160-167 (1999)

[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)

[Publications] Michikawa T.: "Calmodulin mediates calcium-dependent inactivation of the cerebellar type 1 inositol 1,4,5-trisphosphate receptor"Neuron. 23. 799-808 (1999)

[Publications] Furuichi T.: "Intracellular Calucium channels."Calcium as a Cellular Regulator. 49 (1999)

[Publications] Miyawaki A.: "Expression of green fluorescent protein and inositol 1,4,5-triphosphate receptor in xenopus laevis oocytes"Methods in Enzymology. 9 (1999)

[Publications] Mikoshiba K.: "Role of inositol 1,4,5-trisphosphate (IP_3) receptor/Ca^<2+> signaling in cell function"Challenges for Neuroscience in the 21st Century. 26 (1999)

[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)

[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)

[Publications] 竹島浩: "細胞内Ca^<2+>放出チャネル"現代医療. 8 (1999)

[Publications] 古市貞一: "カルシウムチャネル"Clinical Neuroscience(印刷中).

古市貞一理化学研究所, 分子神経形成研究チーム, チームリーダー(研究職) (50219094)