Investigation of membrane binding mechanism of peripheral membrane protein based on solid state NMR using magnetically aligned membrane bilayers at room temperature
| Project/Area Number |
22570126
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | Institute for Molecular Science |
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Principal Investigator |
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| Co-Investigator(Renkei-kenkyūsha) |
TANIO Michikazu 分子科学研究所, 物質分子科学研究領域, 特任助教 (10416662)
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| Project Period (FY) |
2010 – 2012
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| Project Status |
Completed (Fiscal Year 2012)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2011: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2010: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Keywords | 生物科学 / 構造生物化学 / 膜タンパク質 / 固体NMR / 配向試料 / バイセル / 膜表在性 / PLCδ1 / PHドメイン / 開発 / PLC-δ1 |
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| Research Abstract |
Phospholipase C (PLC) binds to phosphatidylinositol 4,5-bisphosphate (PIP_2) in the cell membrane through the pleckstrin homology (PH) domain, and hydrolyzes PIP_2 to produce two second messengers, diacylglycerol and inositol 1,4,5-triphosphate (IP_3), by the catalytic domain. In this study, lipid binding mechanism of human PLC-δ1 PH domain (hPH) was explored. First, the contributions of the α2-helix to the IP_3 binding activity and thermal stability of the hPH domain were investigated by developed approach based on Native-PAGE methods. Then, linked local structural changes of hPH induced by the IP_3 binding were detected by solution NMR. It was found that intra molecular interaction network in hPH controls its ligand binding. Furthermore, through the analyses of solid state NMR, it was also found that hPH induces inhomogeneous structure after binding to a surface of PIP_2 embedded lipid bilayers and implied that hPH is finally inserted to lipid bilayers irreversibly.
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Report
(4 results)
Research Products
(35 results)
