| Project/Area Number |
15GS0312
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| Research Category |
Grant-in-Aid for Creative Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Institution | Osaka University |
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Principal Investigator |
SOBUE Kenji Osaka University, Faculty of Medicine, Professor (20112047)
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| Co-Investigator(Kenkyū-buntansha) |
INUI Makoto Yamaguchi University, Graduate School of Medicine, Professor (70223237)
HAYASHI Kenichiro Osaka University, Graduate School of Medicine, Associate Professor (90238105)
MORITA Tsuyoshi Osaka University, Graduate School of Medicine, Assistant Professor (80403195)
MAYANAGI Taira Osaka University, Graduate School of Medicine, Specially Appointed Assistant Professor (20432544)
柴田 克志 大阪大学, 医学系研究科, 助手 (70296565)
今野 大治郎 大阪大学, 医学系研究科, 助手 (00362715)
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| Project Period (FY) |
2003 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥595,530,000 (Direct Cost: ¥458,100,000、Indirect Cost: ¥137,430,000)
Fiscal Year 2007: ¥127,010,000 (Direct Cost: ¥97,700,000、Indirect Cost: ¥29,310,000)
Fiscal Year 2006: ¥120,250,000 (Direct Cost: ¥92,500,000、Indirect Cost: ¥27,750,000)
Fiscal Year 2005: ¥121,030,000 (Direct Cost: ¥93,100,000、Indirect Cost: ¥27,930,000)
Fiscal Year 2004: ¥114,270,000 (Direct Cost: ¥87,900,000、Indirect Cost: ¥26,370,000)
Fiscal Year 2003: ¥112,970,000 (Direct Cost: ¥86,900,000、Indirect Cost: ¥26,070,000)
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| Keywords | structure biology / synapse / postsvnantic density(PSD) / dopamine receptor / atherosclerosis / vascular smooth muscle cell / LPA receptor / G protein-coupled receptor(GPCR) / 膜貫通型受容体 / GPCR / ドーパミンD1レセプター / PSD蛋白質 / 膜クラスター / LPAレセプター / 膜ターゲティング / G蛋白質共役型受容体(GPCR) / ミリスチン酸化 / 膜蛋白質クラスター / PSD-Zip45 / 神経前駆細胞 / フーリエ変換 / 可塑性 / X線構造解 / G蛋白質共役型受容体 / X線構造解析 / 膜ターゲッティング |
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| Research Abstract |
Transmembrane receptors including a superfamily of G protein-coupled receptors (GPCRs) are essential for all aspects of cell function and also play critical roles for the pathology of a number of diseases. With several exceptions, the three-dimensional ultra-structure of these receptors has not been demonstrated due to the difficultly of their purification in large quantity and their crystallization. In this project, we were developing a novel system for analyzing the three-dimensional ultra-structure of transmembrane receptors. In this system, two-dimensional dusters of transmembrane receptors at paracrystal levels are made in the cell membrane of lived cells by clustering the target receptors through a potently self multimerizable postsynaptic scaffolding protein, PSD-Zip45 (Homer 1c). In order to form a highly ordered two-dimensional transmembrane receptor clusters, we made a drastic modification of PSD-Zip45 (modified PSD-Zip45). As a result, this modified PSD-Zip45 was able to for
… More
m large clusters of dopamine receptor (a typical example of GPCRs) with a diameter of 3-8 um in the cell membrane of lived cells. We then established the purification method for dopamine receptor dusters with a high quality and large quantity. The fourier transformation of electronmicrographs of dopamine receptor duster preparations revealed a high quality at paracrystal levels. This system can be applied to a vast majority of transmembrane receptors. Actually, we also made two-dimensional large dusters of LPA receptor (another example of GPCRs) using our system. Thus, we provide a powerful system for analyzing the structure-function relationship of the transmembrane receptors. For the sake of conventional use, we are now going to try the scale-down of cell culture system for purifying two-dimensional dusters of transmembrane receptors.
In addition to the above project, we further demonstrated the molecular mechanisms underlying neuronal and vascular cell plasticity such as synaptic dynamics mediated by PSD proteins (PSD-Zip45, PSD-Zip70 and others) and onset of atherosclerosis triggered by unsaturated lysophosphatidic adds. Less
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