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1999 Fiscal Year Final Research Report Summary

Structure and function of drug- and ion-extrusion systems in bacteria

Research Project

Project/Area Number 09672230
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biological pharmacy
Research InstitutionOKAYAMA UNIVERSITY

Principal Investigator

TSUCHIYA Tomofusa  Okayama University, Faculty of Pharmaceutical Sciences, Professor, 薬学部, 教授 (80012673)

Co-Investigator(Kenkyū-buntansha) SHIMAMOTO Tadashi  Hiroshima University, Faculty of Applied Biological Science, Associate Professor, 生物生産学部, 助教授 (90187443)
Project Period (FY) 1997 – 1999
KeywordsDrug extrusion system / Ion transport / Pseudomonas aeruginosa / Vibrio parahaemolyticus / Patch clamp
Research Abstract

We investigated drug extrusion system and ion extrusion systems in Vibrio parahaemolyticus, Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus subtilis from structural and functional points of view. Regarding drug extrusion systems, we found several new drug efflux systems, NorM from V. parahaemolyticus, MexXY from P. aeruginosa and EbrAB from B. subtilis. Drug (ethidium) efflux via the NorM was stimulated by Na+. Imposition of inwardly directed Na+ gradient in energy starved cells that were preloaded with ethidium elicited efflux of ethidium. Imposition of inwardly directed ethidium gradient in energy starved cells that were preloaded with Na+ elicited efflux of Na+. These results clearly indicate that drug is extruded from cells in exchange for Na+, namely the NorM is a drug/Na+ antiporter. This is the first example of drug/Na+ antiporter in biological world. Judging from sequence similarity found in databases, it seems that there are some drug/Na+ antiporters in some other bacteria. The MexXY was found to be three-components type multidrug efflux pump. This system is involved in resistance of P. aeruginosa against erythromycin, aminoglycosides and β-lactams. Regarding another efflux systems, ion efflux systems, we found several Na+ efflux systems and analyzed them, Mnh from S. aureus, NhaB from V. parahaemolyticus and NhaG from B. subtilis. These systems extrude Na+ in exchange for H+. The Mnh system is a novel type Na+/H+ antiporter, multisubunit type antiporter. We also analyzed ion efflux from E. coli cells using patch clamp method. This method enabled direct measurement of ion flux in bacterial cells.

  • Research Products

    (28 results)

All Other

All Publications (28 results)

  • [Publications] Rafiqual I. Sarker: "Primary structure and properties of the Na+/glucose symporter (SglS) of Vibrio parahaemolyticus"J. Bacteriol. 179. 1805-1808 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takato Noumi: "Identification and characterization of functional residues in a Na+/H+ antiporter (NhaA) from Escherichia coli by random mutagenesis"J. Biochem.. 121. 661-670 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kei Inaba: "Na+(Li+)/H+ antiporter in Pseudomonas aeruginosa and effect of Li+ on cell growth"Biol. Pharm. Bull.. 20. 621-624 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Teruo Kuroda: "Mutational analysis of amiloride sensitivity in NhaA Na+/H+ antiporter from Vibrio parahaemolyticus"J. Bacteriol. 179. 7600-7602 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kazuyo Kodama: "Preparation and characterization of everted membrane vesicles from cells of Staphylococcus aureus"Biol. Pharm. Bull.. 21. 5-9 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kaori Nozaki: "A new Na+/H+ antiporter, NhaD, of Vibrio parahaemolyticus"Biochim. Biophys. Acta. 1369. 213-220 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Junko Utsugi: "Cloning and sequencing of a novel Na+/H+ antiporter gene from pseudomonas aeruginosa"Biochim. Biophys. Acta. 1398. 330-334 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tomoyuki Mine: "Evidence for chrolamphenico/H+ antiport in Cmr (MdfA) system of Escherichia coli and properties of the antiporter"J. Biochem.. 124. 187-193 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Teruo Kuroda: "Patch-clamp studies on ion pumps of the cytoplasmic membrane of Escherichia coli : Formation, preparation and utilization of giant vacuole-like structures consisting of everted cytoplasmic membrane"J. Biol. Chem.. 273. 16897-16904 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yuji Morita: "NorM, Putative multidrug efflux protein, of Vibrio parahaemolyticus and its homolog in Escherichia coli"Antimicrob. Agents Chemother.. 42. 1778-1782 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hiroki Inoue: "pH-dependent growth retardation of Escherichia coli caused by overproduction of Na+/H+ antiporter"Biol. Pharm. Bull.. 21. 1128-1133 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toshiaki Hiramatsu: "A putative mutisubunit Na+/H+ antiporter from Staphylococcus aureus"J. Bacteriol. 180. 6642-6648 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tomoyuki Mine: "Expression of Escherichia coli of a new multidrug efflux pump, MexXY, from Pseudomonas aeruginosa"Antimicrob. Agents Chemothr.. 43. 415-417 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hiroki Inoue: "Expression of functional Na+/H+ antiporters of helicobacter pylori in antiporter deficient Escherichia coli mutants"FEBS Lett.. 443. 11-16 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Rafiquel I. Sarker, et al.: "Primary structure and properties of the Na+/glucose symporter (SglS) of Vibrio parahaemolyticus"J. Bacteriol.. 179. 1805-1808 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takato Noumi, et al.: "Identification and characterization of functional residues in a Na+/H+ antiporter (NhaA) from Escherichia coli by random mutagenesis"J. Biochem.. 121. 661-670 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kei Inaba, et al.: "NaィイD1+ィエD1(LiィイD1+ィエD1)/HィイD1+ィエD1 antiporter in Pseudomonas aeruginosa and effect of LiィイD1+ィエD1 on cell growth"Biol. Pharm. Bull.. 20. 621-624 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Teuo Kuroda, et al.: "Mutational analysis of amiloride sensitivity in NhaA Na+/H+ antiporter from Vibrio parahaemolyticus"J. Bacteriol.. 179. 7600-7602 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kazuyo Kodama, et al.: "Preparation and characterization of everted membrane vesicles from cells of Saphylococcus aureus"Biol. Pharm. Bull.. 21. 5-9 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kaori Nozaki, et al.: "A new Na+/H+ antiporter, NhaD, of Vibrio parahaemolyticus"Biochem. Biophys. Acta. 1369. 213-220 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Junko Utsugi, et al.: "Cloning and sequencing of a novel Na+/H+ antiporter gene from Pseudomonas aeruginosa"Biochem. Biophys. Acta. 1398. 330-334 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tomoyuki Mine, et al.: "Evidence of chrolamphenicol/H+ antiporter in Cmr (MdfA) system of Escherichia coli and properties of the antiporter"J. Biochem.. 124. 187-193 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Teruo Kuroda, et al.: "Patch-clamp studies on ion pumps of the cytoplasmic membrane of Esherichia coli : Formation, preparation and utilization of giant vacuole-like structures consisting of everted cytoplasmic membrane"J. Biol. Chem.. 273. 16897-16904 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yuji Morita, et al.: "NorM, a putative multidrug efflux protein, of Vibrio parahaemolyticus and its homolog in Escherichia coli"Antimicrob. Agents Chemother. 42. 1778-1782 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hiroki Inoue, et al.: "pH-dependent growth retardation of Escherichia coli caused by oveproduction of Na+/H+ antiporter"Biol. Pharm. Bull.. 21. 1128-1133 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toshiaki Hiramatsu, et al.: "A putative multisubunit Na+/H+ antiporter from Staphylococcus aureus"J. Bacteriol.. 180. 6642-6648 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tomoyuki Mine, et al.: "Expression in Escherichia coli of a new multidrug efflux pump, MexXY, from Pseudomonas aeruginosa"Antimicrob. Agents Chemothr.. 43. 415-417 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hiroki Inoue, et al.: "Expression of functional Na+/H+ antiporters of helicobacter pylori in antiporter deficient Escherichia coli mutants"FEBS Lett.. 443. 11-16 (1999)

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

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Published: 2001-10-23  

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