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2002 Fiscal Year Final Research Report Summary

Towards the dynamic changes of the structure and interaction of ion channels and receptors, -Analysis by Fluorescence Resonance Energy Transfer method-

Research Project

Project/Area Number 12557004
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field General physiology
Research InstitutionTokyo Medical and Dental University graduate School

Principal Investigator

KUBO Yoshihiro  Tokyo Medical and Dental University Graduate School, Department of Physiology, Professor, 大学院・医歯学総合研究科, 教授 (80211887)

Project Period (FY) 2000 – 2002
Keywordsmetabotropic / glutamate / receptor / GFP / FRET
Research Abstract

Ion channels and receptors play important roles in various aspects. The elucidation of structure-function relationship is a physiologically and biophysically interesting theme. In this study, we aimed to analyze the dynamic structural rearrangement of metabotropic glutamate receptor (mGluR1) by FRET (Fluorescence Resonance Energy Transfer) method. We observed so far that mGluR1 can be activated not only by glutamate but also by polyvalent cations, such as Ca^<2+> and Gd^<3+>, and that the sensitivities differ depending on the expression systems including Xenopus oocytes and HEK293 cells. We hypothesized that the difference is due to the difference of phosphorylation, glycosylation, or clustering. As it is known that mGluR1 is composed as homodimers, and that activation occurs by the shift of the equilibrium of inter subunit interaction. We aimed to measure the fluctuation of the inter-subunit interaction of mGluR1 and to estimate the effect of the presence of clustering molecules and polyvalent cations by FRET method. We attached ECFP or EYFP to the carboxy teminal end, and measured FRET between the two subunits. As the recorded FRET efficiency was not high enough, we inserted a new construct with a linker and succeeded in incresing the efficiency. We will continue this study to examine the effect of polyvalent cations and clustering molecules on the inter subunit interaction estimated as FRET efficiency.

  • Research Products

    (26 results)

All Other

All Publications (26 results)

  • [Publications] Miyashita, T., Kubo, Y.: "Extracellular Ca^<2+> sensitivity of mGluR1a associated with persistent glutamate response in transfected CHO cells"Receptors and Channels. 7. 25-40 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miyashita, T., Kubo, Y.: "Extracellular Ca^<2+> sensitivity of mGluR1a induces increase in the basal cAMP level by direct coupling with Gs protein in transfected CHO cells"Receptors and Channels. 7. 77-91 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kubo, Y., Murata, Y.: "Control of rectification and permeation by two distinct sites after the second transmembrane region in kir2.1K^+ channel"Journal of Physiology. 531. 645-660 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Saito, O., Masuho, I., Terakawa, I., Nomoto, S., Asano, T., Kubo, Y.: "RGS8 requires its N-teminus for subcellular localization and acute desensitization of G protein gated K^+ channels"Journal of Biological Chemistry. 276. 5052-5058 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Soom, M., Schonherr, R., Kubo, Y., Kirsch, C., Klinger, R., Heinemanm, S.H.: "Multiple PlP_2 binding sites in Kir2.1 inwardly rectifying potassium channels"FEBS letters. 490. 49-53 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Murata, Y., Okado, H., Kubo, Y.: "Characterization of heteromultimeric G-protein coupled inwardly rectifying potassium channels of the tunicate tadpole with a unique property"Journal of Biological Chemistry. 276. 18529-18539 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Murata, Y., Okado, H., Katsuyama, Y., Okamura, Y., Kubo, Y.: "Primary structure, development expression and functional properties of an inward rectifier K^+ channel of the tunicate"Receptors and Channels. 7. 387-399 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Misaka, T., Miyashita, T., Kubo, Y.: "Primary structure of a dynamin-related mouse mitochondrial GTPase and its distribution in brain, subcellular localization and effect on mitochondrial morphology"Journal of Biological Chemistry. 277. 15834-15842 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ai, T., Fujiwara, Y., Tsuji, K., Otani, H., Nakano, S., Kubo, Y., Horie, M.: "Novel KCNJ2 mutation in familial periodic paralysis with ventricular dysrhythmia"Circulation. 105. 2592-2594 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Saito, O., Murata, Y., Odagiri, M., Itoh, H., Misaka, T., Kubo, Y.: "Altermative splicing of RGS8 gene determines inhibitory function of receptor-type-specific G_9 signaling"Proc. Natl. Acad. Sci USA. 99. 10138-10143 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Murata, Y., Fujiwara, Y., Kubo, Y.: "Idetification of a site involved in the block by extracellular Mg^<2+> and Ba^<2+> as well as permeation of K^+ in Kir21K^+ channel"Journal of Physiology. 544. 665-677 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Fujiwara, Y., Kubo, Y.: "Ser 165 in the second transmembrane region on the Kir2.1 channel determines its susceptibility to blockade by intracellular Mg^<2+>"Journal of General Physiology. 120. 677-692 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Abe, H., Misaka, T., Tateyama, M., Kubo, Y.: "Effects of co-expression with Homer isoforms on the function of metabotropic glutamate receptor 1a"Molecular and Cellular Neuroscience. (in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miyashita, T. and Kubo, Y.: "Extracellular Ca^<2+> sensitivity of mGluR1a associated with persistent glutamate response in transfected CHO cells"Receptors and Channels. 7. 25-40 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Miyashita, T. and Kubo, Y.: "Extracellular Ca^<2+> sensitivity of mGluR1a induces an increase in the basal cAMP level by direct coupling with Gs protein in transfected CHO cells"Receptors and Channels. 7. 77-91 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kubo, Y. and Murata, Y.: "Control of rectification and permeation by two distinct sites after the second transmembrane region in Kir2.1 K^+ channel"Journal of Physiology. 531. 645-660 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Saitoh, O., Masuho, I, Terakawa, I., Nomoto, S., Asano, T. and Kubo, Y: "RGS8 requires its N-terminus for subcellular localization and acute desensitization of G protein gated K^+ channels"Journal of Biological Chemistry. 276. 5052-5058 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Soom, M., Schonherr, R., Kubo, Y., Kirsch, C., Klinger, R. and Heinemann, S. H.: "Multiple PIP_2 binding sites in Kir2.1 inwardly rectifying potassium channels"FEBS letters. 490. 49-53 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Murata, Y., Okado. H. and Kubo, Y.: "Characterization of heteromultimeric G-protein coupled inwardly rectifying potassium channels of the tunicate tadpole with a unique property"Journal of Biological Chemistry. 276. 18529-18539 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Murata, Y., Okado, H., Katsuyama, Y., Okamura, Y. and Kubo, Y.: "Primary structure, developmental expression and functional properties of an inward rectifier K+ channel of the tunicate"Receptors and Channels. 7. 387-399 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Misaka, T., Miyashita, T. and Kubo, Y.: "Primary structure of a dynamin-related mouse mitochondrial GTPase and its distribution in brain, subcellular localization and effect on mitochondrial morphology"Journal of Biological Chemistry. 277. 15834-15842 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ai T., Fujiwara, Y., Tsuji, K., Otani, H., Nakano, S., Kubo, Y. and Horie, M.: "Novel KCNJ2 mutation in familial periodic paralysis with ventricular dysrhythmia"Circulation. 105. 2592-2594 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Saitoh, O., Murata, Y., Odagiri, M., Itoh, M., Itoh, H., Misaka, T. and Kubo, Y.: "Alternative splicing of RGS8 gene determines inhibitory function of receptor-type-specific Gq signaling"Proc. Natl. Acad. Sci. USA. 99. 10138-10143 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Murata,Y., Fujiwara,Y. and Kubo,Y.: "Identification of a site involved in the block by extracellular Mg^<2+> and Ba^<2+> as well as permeation of K^+ in Kir2.1 K^+ channel"Journal of Physiology. 544. 665-677 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Fujiwara, Y. and Kubo, Y.: "Ser 165 in the second transmembrane region of the Kir2.1 channel determines its susceptibility to blockade by intracellular Mg^<2+>"Journal of General Physiology. 120. 677-692 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Abe, H., Misaka, T., Tateyama, M. and Kubo, Y.: "Effects of co-expression with Homer isoforms on the function of metabotropic glutamate receptor1a"Molecular and Cellular Neuroscience. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2004-04-14  

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