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Functional analyzes on an ABC transporter involved in the selective release of outer membrane lipoproteins from membranes

Research Project

Project/Area Number 12660068
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionThe University of Tokyo

Principal Investigator

MATSUYAMA Shin-ichi  Institute of Molecular and Cellular Biosciences, Associate Professor, 分子細胞生物学研究所, 助教授 (50183108)

Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2001: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2000: ¥2,000,000 (Direct Cost: ¥2,000,000)
Keywordslipoprotein / membrane localization / Lol system / ABC transporter / outer membrane / Escherichia coli / ATP / sorting signal / 細胞表層 / ABCトフンスボーター / タンパク質局在化 / リポ蛋白質 / タンパク質輸送 / 局在化 / 相互作用
Research Abstract

We identified a unique ABC transporter, LolCDE, which is responsible for the localization of lipoproteins from the inner to the outer membrane. The ATPase activity of LolCDE was stimulated in an outer membrane lipoprotein-dependent manner, indicating that LolCDE directly recognizes the sorting signal of lipoproteins. The same conclusion was obtained from a genetic analysis on a mutant in which lipoproteins possessing inner membrane sorting signals are localized to the outer membrane. We proposed that the inner membrane sorting signal acts as an avoidance signal on the basis of that inner membrane lipoproteins are not recognized by LolCDE. We showed that N-acylation of the cysteine residue is essential for the Lol system-dependent localization of lipoproteins to the outer membrane. Biochemical analyzes of a mutant LolA protein revealed that LolA directly interacts with LolCDE to transfer lipoproteins. The LolCDE gene as well as lolA and lolB was found to be essential for cell growth of E.coli, and almost all of Gram-negative bacteria have their homologs, suggesting that the membrane localization of lipoproteins by the Lol system is a common feature in Gram-negative bacteria.

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (31 results)

All Other

All Publications (31 results)

  • [Publications] Narita, S.: "A mutation in the membrane subunit of an ABC transporter LolCDE complex causing outer membrane localization of lipoproteins against their inner membrane-specific signals"Mol Microbiol.. (印刷中). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yakushi, T.: "A new ABC transporter mediating the detachment of lipid-modified proteins from membranes"Nature Cell Biology. 2. 212-218 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Masuda, K.: "Elucidation of the function of lipoprotein-sorting signals that determine membrane localization"Proc.Natl.Acad.Sci.USA. 99. 7390-7395 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Fukuda, A.: "Aminoacylation of the N-terminal cysteine is essential for lol-dependent release of lipoproteins from membrane but does not depend on lipoprotein sorting signals"J.Biol.Chem.. 277. 43512-43518 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyamoto, A.: "Dominant negative mutant of a lipoprotein-specific molecular chaperone, LolA, tightly associates with LolCDE"FEBS Lett.. 528. 193-196 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Terada, M.: "Lipoprotein sorting signals evaluated as the LolA-dependent release of lipoproteins from the cytoplasmic membrane of Escherichia coli"J.Biol.Chem.. 276. 47690-47694 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyamoto, A.: "Mutant of LolA, a lipoprotein -specific molecular chaperone of Eschrichia coli, defective in transfer of lipoproteins to LolB"Biochemical and Biophysical research Communications. 287. 1125-1128 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tanaka, K.: "Deletion of lolB encoding an outer membrane lipoprotein, is lethal for Escherichia coli and causes accumulation of lipoprotein localization intermediates in the periplasm"J.Bacteriol.. 183. 6538-6542 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 松山伸一: "大腸菌リポタンパク質の選択的膜局在化機構"実験医学. 21. 874-879 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Narita, S.: "A mutation in the membrane subunit of an ABC transporter LolCDE complex causing outer membrane localization oflipoproteins asainst their inner membrane-specific signals"Mol Microbiol.. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yakushi, T.: "A new ABC transporter mediating the detachment of lipid-modified proteins from membranes"Nature Cell Biol.. 2. 212-218 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Masuda, K.: "Elucidation of the function of lipoprotein-sorting signals that determine membrane localization"Proc. Natl. Acad. Sci. USA. 99. 7390-7395 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Fukuda, A.: "Aminoacylation of the N-terminal cysteine is essential for lol-dependent release of lipoproteins from membrane but does not depend on lipoprotein sorting signals"J. Biol. Chem.. 277. 43512-43518 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyamoto, A.: "Dominant negative mutant of a lipoprotein-specific molecular chaperone, LolA, tightly associates with LolCDE"FEBS Lett.. 528. 193-196 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Terada, M.: "Lipoprotein sorting signals evaluated as the LolA-dependent release of lipoproteins from the cytoplasmic membrane of Escherichia coli"J. Biol. Chem.. 276. 47690-47694 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyamoto, A.: "Mutant of LolA, a lipoprotein -specific molecular chaperone of Escherichia coli, defective in transfer of lipoproteins to LolB"Biochemical and Biophysical Research Communications. 287. 1125-1128 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tanaka, K.: "Deletion of lolB encoding an outer membrane lipoprotein, is lethal for Escherichia coli and causes accumulation of lipoprotein localization intermediates in the periplasm"J. Bacteriol.. 183. 6538-6542 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Matsuyama, S.: "Mechanism of membrane localization of E. coli lipoproteins."Experimental Medicine. 21. 874-879 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 松山伸一: "水に溶けないものが水路を渡る:細菌リポタンパク質の運び屋たち-大腸菌リポタンパク質の選択的膜局在化機構-"実験医学. (印刷中).

    • Related Report
      2002 Annual Research Report
  • [Publications] Fukuda, A.: "Amino-acylation of the N-terminal cysteine is essential for Lol-dependent release of lipoproteins from membranes but does not depend on lipoprotein sorting signals"J. Biol. Chem.. 277. 43512-43518 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Narita, S.: "Disruption of lolCDE encoding an ATP-binding cassette transporter is lethal for Esoherichia coli and prevents the release of lipoproteins from the inner membrane"J. Bacteriol.. 184. 1417-1422 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Masuda, K.: "Elucidation of the function of lipoprotein-sorting signals that determine membrane localization"Proc. Natl. Acad. Sci. USA. 99. 7390-7395 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Miyamoto, A.: "Dominant-negative mutant of a lipoprotein-specific molecular chaperone, LolA, tightly associates with LolCDE"FEBS Lett.. 528. 193-196 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Tanaka, K.: "Deletion of lolB encoding an outer membrane lipoprotein is lethal for Escherichia coli and causes the accumulation of lipoprotein localization intermediates in the periplasm"Journal of Bacteriology. 183. 6538-6542 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Miyamoto, A.: "Mutant of LolA, a lipoprotein-specific molecular chaperone of Escherichia coli, defective in the transfer of lipoproteins to LolB"Biochem. Biophys. Res. Commun.. 287. 1125-1128 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Terada, M.: "Lipoprotein-sorting signals evaluated as the LolA-dependent release of lipoproteins from the cytoplasmic membrane of Escherichia coli"Journal of Biological Chemistry. 276. 47690-47694 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Narita, S.: "Disruption of lolODE encoding an ATP-binding cassette transporter is lethal for Escherichia coli and prevents the release of lipoproteins from the inner membrane"Journal of Bacteriology. 184. 1417-1422 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 徳田 元: "脂質で修飾された蛋白質を膜から遊離させるABCトランスポーター"蛋白質・核酸・酵素. 46. 1221-1227 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 薬師寿治: "グラム陰性細菌におけるリポ蛋白質の生合成機構"日本細菌学会雑誌. 55・3. 517-526 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Hisaharu Yakushi: "A new ABC transporter mediating the detachment of lipid-modified proteins from membranes"Nature Cell Biology. 2・4. 212-218 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Shin-ichi Matsuyama: "ATPase and GTPase activities copurifying with GTP-binding proteins in Escherichia coli"Journal of Molecular Microbiology and Biotechnology. 2・1. 261-263 (2000)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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