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2004 Fiscal Year Final Research Report Summary

Adaptation of cells to high salinity conditions and basic mechanisms of ion transport in biological membranes

Research Project

Project/Area Number 15370054
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Functional biochemistry
Research InstitutionOsaka University

Principal Investigator

KANAZAWA Hiroshi  Osaka University, Graduate School of Science, Professor, 理学研究科, 教授 (50116448)

Co-Investigator(Kenkyū-buntansha) FUKUYAMA Keiichi  Osaka University, Graduate School of Science, Professor, 理学研究科, 教授 (80032283)
NAKAMURA Norihiro  Osaka University, Graduate School of Science, Assistant Professor, 理学研究科, 助手 (90324748)
MITSUI Keiji  Osaka University, Graduate School of Science, Assistant Professor, 理学研究科, 助手 (60379279)
Project Period (FY) 2003 – 2004
KeywordsSalinity torelance / Molecular basis of ion transporters / Na+ / H+ antiporters / genetic and biochemical analyses / bioenergetics of transporters / FRET analyses / Cys scanning mutagenesis / gene engineering
Research Abstract

Intracellular ionic conditions are maintained within a certain range for every ions including H^+ and Na^+. Even for the higher salinity conditions of outside of the cells, cells e remove alkaline cations like Na^+ and Li+ from the inside of cells for keeping homeostasis. This exclusion of toxic alkaline cations from inside of cells is performed by Na^+/H^+ antiporters which exists in most of the cells from bacterial to human. To understand the adaptation mechanisms of cells to the high salinity conditions, molecular mechanism of Na^+/H^+ antiporters should be clarified. In this study, we analyzed molecular structure of Na^+/H^+ antipotrer NhaA from E.coli and H pylori mainly by mutagenic and biochemical studies. As the results, we have shown that essential transmembrane segments for ion transport involves TM4,5,10 and 11 and further shown that essential or important residues in these Tms (J.Biol Chem.(2003)). We also established a method by using FRET(fluorescence resonance energy tra … More nsfer) to detect protein conformational change during ion transport (Manuscript submitted). We isolated a series of mutants with substitution to Cys for each residues in the TM 4,5,10 and 11. Analyses of the mutants in terms of antiporter activities and accessibility of N-ethyl maleimide revealed presense of a putative channel like structure within the NhaA surrounded by the Tms (J.Biol Chem.(2004)). We have cloned new isoforms of human Na^+/H^+ antiporters named NHE8 and 9 and revealed their function and intracellular distribution within different intracellular vesicles (J.Biol.Chem.(2005)). These endomembrane type antiporter function as K^+/H^+ antiporter and regulate endosomal pH. We also revealed a new function of CHP we had found a binding partner of NHE1 previously. We have constructed a knockout cells of gene for CHP and found that NHE does not translocate to the cytoplasmic membrane for the knockout cells, suggesting that CHP is required for intracellular destination of NHE to the cytoplasmic membrane which differs from the previous observation by Pang et al. Less

  • Research Products

    (21 results)

All 2005 2004 2003

All Journal Article (21 results)

  • [Journal Article] Four Na^+/H^+ exchanger isoforms are distributed to Golgi and post-Golgi compartments and are involved in organelle pH regulation.2005

    • Author(s)
      Norihiro Nakamura, Shingo Tanaka, Yoshinori Teko, Hiroshi Kanazawa
    • Journal Title

      J.Biol.Chem. 280

      Pages: 1561-1572

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Characterization of the ion transport activity of budding yeast Na^+/H^+ antiporter, Nha1p, on the secretory vesicles2005

    • Author(s)
      Ryuichi Ohgaki, Norihiro Nakamura, Keiji Mitsui, Hiroshi Kanazawa
    • Journal Title

      Biochem, Biophys.Act-Biomembrane (In press)

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Characterization of the ion transport activity of budding yeast Na^+/H^+ antiporter, Nha1p, on the secretory vesicles2005

    • Author(s)
      Ryuichi Ohgaki, Norihiro Nakamura, Keiji Mitsui, Hiroshi Kanazawa
    • Journal Title

      BBA Biomembrane (In press)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] A novel kinesin-like protein, KIF1Bb3, is involved in the movement of lysosomes to the cell2004

    • Author(s)
      Masafumi Matsushita, Shingo Tanaka, Norihiro Nakamura, Hiroki Inoue, Hiroshi Kanazawa
    • Journal Title

      traffic 5.1

      Pages: 140-151

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] A novel membrane protein capable of binding the Na^+/H^+ antiporter (Nha1p) enhances the salinity-resistant cell growth of Saccharomyces cerevisiae2004

    • Author(s)
      Keiji Mitsui, Fumihiro Ochi, Norihiro Nakamura, Yoshihide Doi, Hiroki Inoue, Hiroshi Kanazawa
    • Journal Title

      J.Biol.Chem. 279

      Pages: 12438-12447

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] A conserved domain in the tail region of the Saccharomyces cerevisiae Na^+/H^+antiporter (Nha1p) plays important roles in localization and salinity-resistant cell-growth2004

    • Author(s)
      Keiji Mitsui, Shinya Kamauchi, Norihiro Nakamura, Hiroki Inoue, Hiroshi Kanazwa
    • Journal Title

      J.Biochem. 135

      Pages: 139-148

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] The Fourth Transmembrane Segment of Helicobacter pylori Na^+/H^+ Antiporter NhaA Faces a Water-filled Channel Required for Ion Transport2004

    • Author(s)
      Naoyuki Kuwabara, Hiroki Inoue, Yumi Tsuboi, Norihiro Nakamura, Hiroshi Kanazawa
    • Journal Title

      J.Biol.Chem. 279

      Pages: 40567-40575

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] A novel kinesin-like protein, KIF1Bb3, is involved in the movement of lysosomes to the cell periphery in non-neuronal cells2004

    • Author(s)
      Masafumi Matsushita, Shingo Tanaka, Norihiro Nakamura, Hiroki Inoue, Hiroshi Kanazawa
    • Journal Title

      Traffic 5

      Pages: 140-151

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] A conserved domain in the tail region of the Saccharomyces cerevisiae Na^+/H^+ antiporter (Nha1p) plays important roles in localization and salinity-resistant cell-growth2004

    • Author(s)
      Keiji Mitsui, Shinya Kamauchi, Norihiro Nakamura, Hiroki Inoue, Hiroshi Kanazawa
    • Journal Title

      J.Biochem. 135

      Pages: 139-148

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Crystal Structure of a Novel Zinc-Binding ATP Sulfurylase from Thermus thermophilus HB82004

    • Author(s)
      Y.Taguchi, M.Sugishima, K.Fukuyama
    • Journal Title

      Biochemistry 43

      Pages: 4111-4118

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Hydroxylamine and Hydrazine Bind Directly to the Heme Iron of the Heme-Heme Oxygenase-1 Complex.2004

    • Author(s)
      H.Sakamoto, Y.Higashimoto, S.Hayashi, M.Sugishima, K.Fukuyama, G.Palmer, M.Noguchi, J.Inorg
    • Journal Title

      Biochem. 98

      Pages: 1223-1228

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] CO-trapping Site in Heme Oxygenase Revealed by Photolysis of its CO-bound Heme Complex : Mechanism of Escaping from Product Inhibition2004

    • Author(s)
      M.Sugishima, H.Sakamoto, M.Noguchi, K.Fukuyama
    • Journal Title

      J.Mol.Biol. 341

      Pages: 7-13

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Structure and function of plant-type ferredoxins.2004

    • Author(s)
      K.Fukuyama
    • Journal Title

      Photosynth.Res. 81

      Pages: 289-301

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Calcineurin homologous protein isoform 2 (CHP2), Na+/H+ exchangers- biding protein, is expressed in intestinal epithelium2003

    • Author(s)
      H.Inoue, Y.Nakamura, M.Nagita, I.Takai, M.Masuda, N.Nakamura, H.Kanazawa
    • Journal Title

      Bull.Pharm.Bull. 26

      Pages: 148-155

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Identification of membrane domains of Na^+/H^+ antiporter (NhaA) from Helicobacter pylori required for ion transport and pH sensing2003

    • Author(s)
      Yumi Tsuboi, Hiroki Inoue, Norihiro Nakamuara, Hiroshi Kanazawa
    • Journal Title

      J.Biol.Chem. 278

      Pages: 21467-21473

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] The apoptosis-inducing protein kinase DRAK2 is inhibited in a calcium-dependent manner by the calcium-binding protein CHP.2003

    • Author(s)
      Hiroshi Kuwahara, Junichi Kamei, Norihiro Nakamura, Miho Matsumoto, Hiroki, Inoue
    • Journal Title

      J.Biochem. 134

      Pages: 245-250

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Two nuclear export signals specify the cytoplasmic localization of calcineurin B homologous protein 12003

    • Author(s)
      Mana Nagaita, Hiroki Inoue, Norihiro Nakamura, Hiroshi Kanazawa
    • Journal Title

      J.Biochem. 134

      Pages: 919-925

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Calcineurin homologous protein isoform 2 (CHP2), Na+/H+ exchangers-binding protein, is expressed in intestinal epithelium2003

    • Author(s)
      H.Inoue, Y.Nakamura, M.Nagita, T.Takai, M.Masuda, N.Nakamura, H.Kanazawa
    • Journal Title

      Bull.Pharm.Bull. 26

      Pages: 148-155

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Identification of membrane domains of Na^+/H^+ antiporter (NhaA) from Helicobacter pylori required for ion transport and pH sensing2003

    • Author(s)
      Yumi Tsuboi, Hiroki Inoue, Norihiro Nakamura, Hiroshi Kanazawa
    • Journal Title

      J.Biol.Chem. 278

      Pages: 21467-21473

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] The apoptosis-inducing protein kinase DRAK2 is inhibited in a calcium-dependent manner by the calcium-binding protein CHP2003

    • Author(s)
      Hiroshi Kuwahara, Junichi Kamei, Norihiro Nakamura, Miho Matsumoto, Hiroski, Inoue, Hiroshi Kanazawa
    • Journal Title

      J.Biochem. 134

      Pages: 245-250

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Two nuclear export signals specify the cytoplasmic localization of calcineurin B homologous protein 12003

    • Author(s)
      Mana Nagaita, Hiroki Inoue, Norihiro Nakamura, Hiroshi Kanazawa
    • Journal Title

      J.Biohcem. 134

      Pages: 919-925

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

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Published: 2006-07-11  

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