| Project/Area Number |
10308029
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| Research Category |
Grant-in-Aid for Scientific Research (A).
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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 | Osaka University |
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Principal Investigator |
YAMAGUCHI Akihito Industrial and Scientific Research Institute (ISIR), Osaka University Cell Membrane Biology Professor, 産業科学研究所, 教授 (60114336)
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| Co-Investigator(Kenkyū-buntansha) |
NADA Shigeyuki ISIR, Osaka University, Cell Membrane Biology, Research Associate, 産業科学研究所, 助手 (50291448)
MURAKAMI Satoshi ISIR, Osaka University, Cell Membrane Biology, Research Associate, 産業科学研究所, 助手 (30300966)
染谷 雄一 国立感染症研究所, 研究員 (50283809)
森山 芳則 岡山大学薬学部, 教授 (10150658)
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥31,800,000 (Direct Cost: ¥31,800,000)
Fiscal Year 2000: ¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1999: ¥9,600,000 (Direct Cost: ¥9,600,000)
Fiscal Year 1998: ¥15,600,000 (Direct Cost: ¥15,600,000)
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| Keywords | xenobiotic exporter / drug resistance / site-directed mutagenesis / efflux / exporter / tetracycline / X-ray crystallography / post genome / 多剤耐性 / アンチポーター / cMOAT / Acr / ABCトランスポーター / Tet(B) |
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| Research Abstract |
(1) We constructed the complete set of cysteine-scanning mutants of tetracycline/H+ antiporter TetA (B) and investigated the detailed topology by means of site-directed chemical modification and oxidative disulfide cross-linking formation of double Cys mutants. As a result, it was revealed that TetA (B) forms a circular form and involves a water-filled channel at the center of the molecule with a permeability barrier in the middle of the membrane.
(2) We established the method for measuring the drug efflux mediated by MRP2 by means of fluorescent drugs. Then, we determined the transmembrane basic residues essential for its transport function and the binding of the inhibitor by means of site-directed mutagenesis.
(3) We first succeeded to obtain crystals of xenobiotic exporters. Now, we are undergoing the X-ray crystallographic analysis using SPring8.
(4) We found a novel possible exporter for sphingosine-1-phosphate from platelete and its complete cDNA was cloned and expressed in a cell line.
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