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Studies in membrane topology and function of inner membrane component, MexA,of antibiotic extrusion pump of Pseudomonas aeruginosa

Research Project

Project/Area Number 09670298
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Bacteriology (including Mycology)
Research InstitutionTokai University

Principal Investigator

YONEYAMA Hiroshi  Tokai University, School of Medicine, Assistant Professor, 医学部, 講師 (10220774)

Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1998: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1997: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsPseudomonas aeruginosa / multidrug resistance / extrusion pump / inner membrane protein / outer membrane protein / antibiotics / 他剤耐性
Research Abstract

Pseudomonas aeruginosa, a main causative agent of opportunistic infections, shows intrinsic and multiantibiotic resistance. The major reason for the antibiotic resistance in this bacterium appeared to be the presence of an energy dependent antibiotic extrusion pump (MexAB-OprM pump) encoded in the mex operon (mexA-mexl3-oprM). To get insight into the mode of antibiotic extrusion by the Mex pump, it is essential to study on the structure and function of each subunit of the pump. Therefore, I analyzed the membrane topology and localization of MexA, a member of membrane fusion protein family, which is thought to link inner membrane pump, MexB, to an outer membrane channel, OprM.I employed the gene fusion technique to analyze the membrane topology of MexA using signal sequenceless beta-lactamase as a reporter and got 78 in-frame MexA-beta--lactamase fusions. All transformants harbored each fusion gene showed ampicillin resistance, indicating that almost all regions of MexA beyond the signal sequence reside in the periplasm. MexA was also found to be modified by lipid(s) at its cysteine residue in the consensus sequence for lipoproteins.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Yoneyama et al.: "Subunit swapping in the Mex-extrusion pumps in Pseudomonas aeruginosa." Biochem.Biophys.Res.Commun.244. 898-902 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yoshihara et al.: "Identification of the catalytic triad of the protein D2 protease in Pseudomonas aeruginosa." Biochem.Biophys.Res.Commun.247. 142-145 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yoneyama et al.: "The role of mex-gene products in antibiotic extrusion in Pseudomonas aeruginosa." Biochem.Biophys.Res.Commun.233. 611-618 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ocaktan et al.: "Use of fluorescence probes to monitor function of the subunit proteins of the MexA- MexB- OporM drug extrusion machinery in Pseudomonas aeruginosa." J.Biol.Chem.272. 21964-21969 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kitahara et al.: "Antibiotic diffusion pathways in the outer membrane of Pseudomonas aeruginosa." Biochem.Biophys.Res.Commun.238. 457-461 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nakae et al.: "Multiantibiotic resistance caused by active drug extrusion in hospital pathogens." J.Infect.Chemother.3. 173-183 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yoneyama, H., Ocaktan, A., Tsuda, M.and Nakae, T.: "The role of mex-gene products in antibiotic extrusion in Pseudomonas aeruginosa." Biochem.Biophys.Res.Commun.233. 611-618 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ocaktan, A., Yoneyama, H.and Nakae, T.: "Use of fluorescence probes to monitor function of the subunit proteins of the MexA-MexB-OporM drug extrusion machinery in Pseudomonas aeruginosa." J.Biol.Chem.272. 21964-21969 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kitahara, T., Yoneyama, H.and Nakae, T.: "Antibiotic diffusion pathways in the outer membrane of Pseudomonas aeruginosa." Biochem.Biophys.Res.Commun.238. 457-461 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nakae, T., Yoshihara, E.and Yoneyama, H.: "Multiantibiotic resistance caused by active drug extrusion in hospital pathogens." J.Infect.Chemother.3. 173-183 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yoneyama, H., Ocaktan, A., Gotoh, N., Nishino, T.and Nakae, T.: "Subunit swapping in the Mex-extrusion pumps in Pseudomonas aeruginosa." Biochem.Biophys.Res.Commun.244. 898-902 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yoshihara, E., Yoneyama, H., Ono, T.and Nakae, T.: "Identification of the catalytictriad of the protein D2 protease in Pseudomonas aeruginosa." Biochem.Biophys.Res.Commun.247. 142-145 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yoneyama et al.: "Subunit swapping in the Mex-extrusion pumps in Pseudomonas aeruginosa." Biochem.Biophys.Res.Commun.244. 898-902 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yoshihara et al.: "Identification of the catalytic triad of the protein D2 protease in Pseudomonas aeruginosa." Biochem.Biophys.Res.Commun.247. 142-145 (1998)

    • Related Report
      1998 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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