2000 Fiscal Year Final Research Report Summary
Molecular Basis and Physiological Roles of Xenobiotic Exporters
| 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)
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| Project Period (FY) |
1998 – 2000
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| Keywords | xenobiotic exporter / drug resistance / site-directed mutagenesis / efflux / exporter / tetracycline / X-ray crystallography / post genome |
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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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[Publications] Kimura-Someya,T.,Iwaki,S.,Konishi,S.,Tamura,N.,Kubo,Y.and Yamaguchi,A.: "Cysteine-scanning Mutagenesis around Transmembrane Segemnts 1 and 11 and Their Flanking Loop Regions of Tn10-encoded Metal-Tetracycline/H^+ Antiporter."J.Biol.Chem. 275. 18692-18697 (2000)
Description
「研究成果報告書概要(和文)」より
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[Publications] Tamura,N.,Konishi,S.,Iwaki,S.,Kimura-Someya,T.,Nada,S.,and Yamaguchi,A.: "Complete Cysteine-Scanning Mutagenesis and Site-Directed Chemical Modification of the Tn10-Encoded Metal-Tetracycline/H^+ Antiporter."J.Biol.Chem. 276(in press). (2001)
Description
「研究成果報告書概要(和文)」より
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[Publications] Kimura-Someya, T., Iwaki, S., Konishi, S., Tamura, N., Kubo, Y.and Yamaguchi, A.: "Cysteine-scanning Mutagenesis around Transmembrane Segemnts 1 and 11 and Their Flanking Loop Regions of Tn10-encoded Metal-Tetracycline/H^+ Antiporter."J.Biol.Chem.. 275. 18692-18697 (2000)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Tamura, N., Konishi, S., Iwaki, S., Kimura-Someya, T., Nada, S., and Yamaguchi, A.: "Complete Cysteine-Scanning Mutagenesis and Site-Directed Chemical Modification of the Tn10-Encoded Metal-Tetracycline/H^+ Antiporter."J.Biol.Chem.. 276(in press). (2001)
Description
「研究成果報告書概要(欧文)」より
