1993 Fiscal Year Final Research Report Summary
Development of Highly Sensitive Bilayr Lipid Membrane Sensors Based on Membrane Proteins
| Project/Area Number |
04453156
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Physical pharmacy
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| Research Institution | University of Tokyo (1993)
Hokkaido University (1992) |
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Principal Investigator |
ODASHIMA Kazunori University of Tokyo Graduate School of Pharmaceutical Sciences, Associate Professor, 大学院・薬学系研究科, 助教授 (30152507)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAYAMA Hitoshi Kumamoto University Faculty of Pharmaceutical Sciences, Professor, 薬学部, 教授 (70088863)
TOHDA Koji University of Tokyo School of Science, Assistant Professor, 大学院・理学系研究科, 助手 (60212065)
SUGAWARA Masao University of Tokyo School of Science, Associate Professor, 大学院・理学系研究科, 助教授 (50002176)
UMEZAWA Yoshio University of Tokyo School of Science, Professor, 大学院・理学系研究科, 教授 (80011724)
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| Project Period (FY) |
1992 – 1993
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| Keywords | Biosensor / Ion Channel Protein / Bilayr Lipid Membrane / Glutamate Receptor Ion Channel / NMDA Subtype / Voltage-sensitive Ion Channel / Channel Current / Agonist Selectivity |
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| Research Abstract |
Fundamental studies aiming at generalization of bilayr lipid membrane sensors, which exploit remarkable signal amplification function of ion channel proteins for highly sensitive sensing of target compounds, were carried out and the following results were obtained.
1.Glutamate receptor ion channel protein (GluR), purified from postsynaptic membranes of rat brain, was incorporated into a planar bilayr lipid membrane, and its agonist selectivity was evaluated for several representative agonists under the conditions in which only the NMDA subtype was active. The agonist selectivity based on the total ion permeated through the channels (Ca^<2+> and Na^+) was lower than that of the binding assay, although the order of the selectivity was the same for the two systems. In addition, the agonist selectivity based on the permeation of Ca^<2+> ions was evaluated by thin layr potentiometry using liposomes incorporated with the GluR proteins. A selectivity similar to that for planar bilayr lipid mem
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branes was observed for the NMDA subtype. The present approach based on the transmembrane signals from the GluR proteins (total ions permeated through the membrane) affords a new method for evaluating a biocompatible selectivity, which is different from the binding affinity used so far to evaluate the agonist selectivities of channel proteins.
2.Fabrication of a hybrid-type bilayr lipid membrane, incorporated with voltage-sensitive sodium channel proteins (from electric organ of electric eel) with artificial receptor molecules capable of generating guest-induced membrane potential changes, was attempted. Planar bilayr lipid membranes incorporated with the above ion channel protein and valinomycin were prepared. Although the incorporated channel proteins kept ion channel activity, addition of K^+ ion in low concentrations caused a marked decrease in the membrane resistance, indicating the necessity of reinvestigation of the experimental system to make it able to measure the changes in channel currents involving membrane potential changes. Less
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[Publications] E.K.Michaelis, M.L.Michaelis, K.N.Kumar, N.Tilakaratne, D.B.Joseph, P.S.Johnson, K.K.Babcock, G.L.Aistrup, R.L.Schowen, H.Minami, M.Sugawara, K.Odashima, and Y.Umezawa: ""Purification, Reconstitution, and Cloning of an NMDA Receptor-Ion Channel Complex from Rat Brain Synatic Membranes : Implications for Neurobiological Changes in Alcoholism"" Ann.New York Acad.Sci.654. 7-18 (1992)
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「研究成果報告書概要(欧文)」より
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[Publications] M.Sugawara, N.Sugao, Y.Umezawa, Y.Adachi, K.Taniguchi, H.Minami, M.Uto, K.Odashima, E.K.Michaelis, and T.Kuwana: ""Transmembrane Signalling by a Transport or Receptor Ion Channel Protein Embedded in Planar Bilayr Lipid Membranes : Detection of Bioactive Substances"" Proceedings of the Fifth International Symposium on Redox Mechanisms and Interfacial Properties of Molecules of Biological Importance, ed.by F.A.Schultz and I.Taniguchi, The Electrochemical Society, Pennington, NJ,U.S.A. 268-279 (1993)
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「研究成果報告書概要(欧文)」より
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