Dynamic structural rearrangements of ion channels and receptors dependent on expression density.
| Project/Area Number |
16390052
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General physiology
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| Research Institution | National Institute for Physiological Sciences |
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Principal Investigator |
KUBO Yoshihiro National Institute for Physiological Sciences, Department of Molecular Physiology, Professor, 分子生理研究系, 教授 (80211887)
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| Co-Investigator(Kenkyū-buntansha) |
SATO Chikara National Institute of Advanced Science and Technology, Neuroscience Research Institute, Group Leader, 脳神経情報研究部門, グループ長 (00357146)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥13,800,000 (Direct Cost: ¥13,800,000)
Fiscal Year 2005: ¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 2004: ¥7,400,000 (Direct Cost: ¥7,400,000)
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| Keywords | ATP receptor / P2X / single particle structure analysis / phospholipids / structural rearrangement / FRET / structure-function relationship / functional ragulation / 膜電位依存性 / ゲート機構 / ATP受容体チャネル / P2X2 / バキュウロウイルス / Sf9細胞 |
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| Research Abstract |
Our aim of research is to elucidate the mechanisms of the environment dependent changes of ion channels and receptor function and to know their dynamic structural rearrangements. During the period of this grant, we have conducted four specific researches in the following. [1]We, for the first time, observed that the pore properties and the ligand sensitivity of ATP receptor channel P2X2 change depending on "the open channel density on the membrane" probably by mutual interaction between them. We also uncovered by systematic mutagenesis study that Ile328 in the second transmembrane region is critical for the expression density dependent changes. [2]We purified recombinant protein of P2X2 by baculo virus - Sf9 cell expression system, and carried out stoichiometry analyses by anchoring the recombinant proteins with glutaraldehyde. We observed that P2X2 is composed of three subunits. We also performed single particle structure analysis using the recombinant proteins, and confirmed that P2X2 is a trimeric protein which shows an inverted pyramid like structure. [3]As a possible mechanism of the density dependent changes of P2X2, we analyzed phospholipids dependency. We observed that the P2X2 function depends not on PIP2 but on PIP3, and that the dilated pore state is directly linked to the desensitized state. [4]We analyzed the structural rearrangements of the metabotropic glutamate receptor by FRET technique, and observed that the relative allocation of the dimeric subunits changes upon glutamate application. We also observed that the type of coupling G proteins of mGluR1 changes depending on the type of ligand, showing that mGluR1 is not a simple on-/off- switch but a multiple path regulator.
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Report
(3 results)
Research Products
(27 results)
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[Journal Article] OPA1 expression in the normal rat retina and optic nerve.2005
Author(s)
Ju, W.K., Misaka, T., Kushnareva, Y., Nakagomi, S, Agarwal, N., Kubo, Y.et al.
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Journal Title
Journal of Comparative Neurology 488
Pages: 1-10
Related Report
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