Molecular mechanisms underlying the membrane sorting of bacterial lipoproteins

• Project/Area Number: 19380046
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: The University of Tokyo
• Principal Investigator: TOKUDA Hajime The University of Tokyo, 分子細胞生物学研究所, 教授 (40125943)
• Project Period (FY): 2007 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥18,850,000 (Direct Cost: ¥14,500,000、Indirect Cost: ¥4,350,000); Fiscal Year 2009: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2008: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000); Fiscal Year 2007: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
• Keywords: 膜局在化 / ABCトランスポーター / 分子シャペロン / ペリプラズム / リポ蛋白質 / 光架橋反応 / LolA / LolB / LolCDE / 選別シグナル / 大腸菌

Research Abstract

Lipoproteins possessing the lipid-attached cysteine residue at the N-terminus are widely distributed among bacteria and anchored to membranes through the N-terminal lipids. Lipoproteins are involved in various functions present in cell surfaces and localized in either the inner or outer membrane of Gram-negative bacteria depending on their sorting signals. We have reveled that five Lol factors are involved in the membrane sorting of lipoproteins. The mechanisms underlying the specific membrane sorting of lipoproteins by Lol proteins are examined in this study.

Research Products

• [Journal Article] A novel complete reconstitution system for membrane integration of the simplest membrane protein (2010)
  • Author(s): Nishiyama, K., Maeda, M., Abe, M., Kanamori, T., Shimamoto, K., Kusumoto, S., Ueda, T., Tokuda, H.
  • Journal Title: Biochem.Biophys.Res.Commun. 394 Pages: 733-736
  • NAID: 120002439245
• [Journal Article] Biogenesis of outer membranes in Gram-negative bacteria (2009)
  • Author(s): Tokuda, H.
  • Journal Title: Biosci.Biotechnol.Biochem. 73 Pages: 465-473
  • NAID: 10027537908
• [Journal Article] Model of mouth-to-mouth transfer of bacterial lipoproteins through inner membrane LolC, periplasmic LolA, and outer membrane LolB. (2009)
  • Author(s): Okuda, S., Tokuda, H.
  • Journal Title: Proc.Natl.Acad.Sci., USA 106 Pages: 5877-5882
• [Journal Article] Dissection of the LolB function ; lipoprotein binding, membrane targeting, and incorporation of lipoproteins into lipid bilayers (2009)
  • Author(s): Tsukahara, J., Mukaiyama, K., Okuda, S., Narita, S., Tokuda, H.
  • Journal Title: FEBS J. 276 Pages: 4496-4504
• [Journal Article] Biochemical characterization of an ABC transporter LptBFGC complex required for the outer membrane sorting of lipopolysaccharides (2009)
  • Author(s): Narita, S., Tokuda, H.
  • Journal Title: FEBS Lett. 583 Pages: 2160-2164
• [Journal Article] Structural investigation of the interaction between LolA and LolB using NMR. (2009)
  • Author(s): Nakada, S., Sakakura, M., Takahashi, H., Okuda, S., Tokuda, H., Shimada, I.
  • Journal Title: J.Biol.Chem. 284 Pages: 24634-24643
• [Journal Article] Membrane topology and functional importance of the periplasmic region of ABC transporter LolCDE (2009)
  • Author(s): Yasuda, M., Iguchi-Yokoyama, A., Matsuyama, S., Tokuda, H., Narita, S.
  • Journal Title: Biosci.Biotechnol.Biochem. 73 Pages: 2310-2316
  • NAID: 10027547063
• [Journal Article] Real time analysis of lipoprotein transfer from LolA to LolB by means of surface plasmon resonance (2009)
  • Author(s): Tsukahara, J., Narita, S., Tokuda, H.
  • Journal Title: FEBS Lett. 583 Pages: 2987-2990
• [Journal Article] Development of a functional in vitro integration system for an integral membrane protein, SecG. (2009)
  • Author(s): Nishiyama, K., Tokuda, H.
  • Journal Title: Biochem.Biophys.Res.Commun. 390 Pages: 920-924
• [Journal Article] Real time analysis of lipoprotein transfer from LolA to LolB by means of surface plasmon resonance. (2009)
  • Author(s): Tsukahara, J.
  • Journal Title: FEBS Lett. 583 Pages: 2987-2990
  • Peer Reviewed
• [Journal Article] Membrane Topology and Functional Importance of the Periplasmic Region of ABC Transporter LolCDE. (2009)
  • Author(s): Yasuda M.
  • Journal Title: Biosci.Biotechnol.Biochem. 73 Pages: 2310-2316
  • Peer Reviewed
• [Journal Article] Dissection of the LolB function ; lipoprotein binding, membrane targeting, and incorporation of lipoproteins into lipid bilayers. (2009)
  • Author(s): Tsukahara, J.
  • Journal Title: FEBS J. 276 Pages: 4496-4504
  • Peer Reviewed
• [Journal Article] Structural investigation of the interaction between LolA and LolB using NMR. (2009)
  • Author(s): Nakada, S.
  • Journal Title: J.Biol.Chem. 284 Pages: 24634-24643
  • Peer Reviewed
• [Journal Article] Model of mouth-to-mouth transfer of bacterial lipoproteins through inner membrane LolC, periplasmic LolA, and outermembrane LolB. (2009)
  • Author(s): Okuda, S.
  • Journal Title: Proc.Natl.Acad.Sci.,USA 106 Pages: 5877-5882
  • Peer Reviewed
• [Journal Article] Molecular events involved in a single cycle of ligand transfer from an ATP-binding cassette transporter, LolCDE, to a molecular chaperone, LolA. (2008)
  • Author(s): Taniguchi, N., Tokuda, H.
  • Journal Title: J.Biol.Chem. 283 Pages: 8538-8544
• [Journal Article] A short helix in the C-terminal region of LolA is important for the specific membrane localization of lipoproteins (2008)
  • Author(s): Okuda, S., Watanabe, S., Tokuda, H.
  • Journal Title: FEBS Lett. 582 Pages: 2247-2251
• [Journal Article] Diacylglycerol specifically blocks spontaneous integration of membrane proteins and allows detection of a factor-assisted integration (2008)
  • Author(s): Kawashima, Y., Miyazaki, E., Muller, M., Tokuda, H., Nishiyama, K.
  • Journal Title: J.Biol.Chem. 283 Pages: 24489-24496
• [Journal Article] Opening and closing of the hydrophobic cavity of LolA coupled to lipoprotein binding and release (2008)
  • Author(s): Oguchi, Y., Takeda, K., Watanabe, S., Yokota, N., Miki, K., Tokuda, H.
  • Journal Title: J.Biol.Chem. 283 Pages: 25414-25420
• [Journal Article] Introduction of a lethal redox switch that controls the opening and closing of the hydrophobic cavity in LolA. (2008)
  • Author(s): Watanabe, S., Oguchi, Y., Takeda, K., Miki, K., Tokuda, H.
  • Journal Title: J.Biol.Chem. 283 Pages: 25421-25427
• [Journal Article] Molecular Events Involved in a Single Cycle of Ligand Transfer from an ATP Binding Cassette Transporter, LolCDE, to a Molecular Chaperone, LolA. (2008)
  • Author(s): Taniguchi, N.
  • Journal Title: J. Biol. Chem. 283 Pages: 8538-8544
  • Peer Reviewed
• [Journal Article] A short helix in the C-terminalregion of LolA is important for the specific membrane localization of lipoproteins. (2008)
  • Author(s): Okuda, S.
  • Journal Title: FEBS Lett 582 Pages: 2247-2251
  • Peer Reviewed
• [Journal Article] Introduction of a lethal redox switch that controls the opening and closing of the hydrophobic cavity in LoLA. (2008)
  • Author(s): Watanabe, S.
  • Journal Title: J. Biol. Chem. 283 Pages: 25421-25427
  • Peer Reviewed
• [Journal Article] Opening and closing of the hydrophobic cavity of LolA coupled to lipo protein binding and release. (2008)
  • Author(s): Oguchi, Y.
  • Journal Title: J. Biol. Chem. 283 Pages: 25414-25420
  • Peer Reviewed
• [Journal Article] A new screening method to identify inhibitors of Lol (localization of lipoproteins) system, a novel antibacterial target (2007)
  • Author(s): Ito, H., Ura, A., Oyamada, Y., Yoshida, H., Yamagishi, J., Narita, S., Matsuyama, S., Tokuda, H.
  • Journal Title: Microbiol.Immunol. 51 Pages: 263-270
  • NAID: 10020280998
• [Journal Article] Amino acids at positions 3 and 4 determine the membrane specificity of Pseudomonas aeruginsoa lipoproteins (2007)
  • Author(s): Narita, S., Tokuda, H.
  • Journal Title: J.Biol.Chem. 282 Pages: 13372-13378
• [Journal Article] Characterization of the Pseudomonas aeruginosa Lol system as a lipoprotein sorting mechanism (2007)
  • Author(s): Tanaka, S., Narita, S., Tokuda, H.
  • Journal Title: J.Biol.Chem. 282 Pages: 13379-13384
• [Journal Article] Diverse effects of phospholipids on lipoprotein sorting and ATP hydrolysis by the ABC transporter LolCDE complex. (2007)
  • Author(s): Miyamoto, S., Tokuda, H.
  • Journal Title: Biochim.Biophys.Acta-Biomembranes 1768 Pages: 1848-1854
• [Journal Article] Complete reconstitution of an ATP-binding cassette transporter LolCDE complex from separately isolated subunits (2007)
  • Author(s): Kanamaru, K., Miyamoto, S., Narita, S., Tokuda, H.
  • Journal Title: FEBS J. 274 Pages: 3034-3043
• [Journal Article] Large-scale preparation of the homogeneous LolA-lipoprotein complex and efficient in vitro transfer of lipoproteins to the outer membrane in a LolB-dependent manner (2007)
  • Author(s): Watanabe, S., Oguchi, Y., Yokota, N., Tokuda, H.
  • Journal Title: Protein Sci. 16 Pages: 2741-2749
• [Journal Article] A new screening method to identify inhibitors of Lol(localization of lipoproteins)system,anovel antibacterial target (2007)
  • Author(s): Ito, H.
  • Journal Title: Microbiol.Immunol 51 Pages: 263-270
  • Peer Reviewed
• [Journal Article] Amino acids at positions 3 and 4 determine the membrane specificity of Pseudomonas aeruginosa lipoproteins (2007)
  • Author(s): Narita, S.
  • Journal Title: J.Biol.Chem 282 Pages: 13372-13378
  • Peer Reviewed
• [Journal Article] Characterization of the Pseudomonas aeruginosa Lol system as a lipoprotein sorting mechanism (2007)
  • Author(s): Tanaka, S.
  • Journal Title: J.Biol.Chem 282 Pages: 13379-13384
  • Peer Reviewed
• [Journal Article] Diverse effects of phospholipids on lipoprotei sorting and ATP hydrolysis by the ABC transporter LolCDE complex (2007)
  • Author(s): Miyamoto, S.
  • Journal Title: Biochim.Biophys.Acta 1768 Pages: 1848-1854
  • Peer Reviewed
• [Journal Article] Complete reconstitution of an ATP-binding cassette transporter LolCDE complex from separately isolated subunits (2007)
  • Author(s): Karlamaru, K.
  • Journal Title: FEBS J 274 Pages: 3034-3043
  • Peer Reviewed
• [Journal Article] The function of the ABC transporter LolCDE in protein transport to the outer membrane of E.coli (2007)
  • Author(s): Narita, S.
  • Journal Title: The Enzymes 25 Pages: 149-172
  • Peer Reviewed
• [Journal Article] Structural studies of the Cpx pathway activator NlpE on the outer membrane of Escherichia coli (2007)
  • Author(s): Hirano, Y.
  • Journal Title: Structure 15 Pages: 963-976
  • Peer Reviewed
• [Journal Article] Backbone resonance assignment for the outer membrane lipoprotein receptor LolB from Escherichia coli (2007)
  • Author(s): Nakada, S.
  • Journal Title: Biomol.NMR Assign 1 Pages: 121-123
  • Peer Reviewed
• [Journal Article] Topology inversion of SecG is essential for the cytosolic SecA-dependent stimulation of protein translocation (2007)
  • Author(s): Sugai, R.
  • Journal Title: J.Biol.Chem 282 Pages: 29540-29548
  • Peer Reviewed
• [Journal Article] Large-scale preparation of the homogeneous LolA-lipoprotein complex and efficient in vitro transfer of lipoproteins to the outer membrane in a LolB-deperldent manner (2007)
  • Author(s): Watanabe, S.
  • Journal Title: Protein Sci 16 Pages: 2741-2749
  • Peer Reviewed
• [Presentation] Sorting of lipoproteins to the outer membrane of Gram-negative bacteria (2010)
  • Author(s): Tokuda, H.
  • Organizer: 3rd FEBS Special Meetig "ATP-Binding Cassette Proteins : From Multidrug Resistance to Genetic Diseases"
  • Place of Presentation: Innsbruck(Austria)
  • Year and Date: 2010-03-04
• [Presentation] Sorting of lipoproteins to the outer membrane of Gram-negative bacteria (2010)
  • Author(s): Tokuda, H.
  • Organizer: Final Keynote Address, FEBS Special Meeting "ATP-binding cassette proteins : from multidrug resistance to genetic diseases"
  • Place of Presentation: Innsbruck, Austria
• [Presentation] in vivo光架橋法によるLol因子間相互作用の解析 (2008)
  • Author(s): 奥田傑
  • Organizer: 21世紀大腸菌研究会
  • Place of Presentation: 静岡国民年金健康センター
  • Year and Date: 2008-07-28
• [Presentation] 部位特異的光架橋法によるLolA-LolB間相互作用のin vivo解析 (2008)
  • Author(s): 奥田 傑
  • Organizer: 日本農芸化学会2008年度大会
  • Place of Presentation: 名城大学(名古屋)
  • Year and Date: 2008-03-27
• [Presentation] 細菌リポタンパク質の選別と外膜局在化機構 (2008)
  • Author(s): 徳田 元
  • Organizer: 第81回日本細菌学会総会
  • Place of Presentation: 国立京都国際会館(京都)
  • Year and Date: 2008-03-24
• [Presentation] Molecular events involved in a single cycle of ligand transfer from an ATP-binding cassette transporter LolCDE to a molecular chaperone LoIA. (2008)
  • Author(s): Tokuda, H.
  • Organizer: 2nd FEBS Special Meeting"ATP-Binding Cassete Proteins"
  • Place of Presentation: Hotel Crauer Bar(Innsbruck,Austria)
  • Year and Date: 2008-03-02
• [Presentation] Molecular events involved in a single cycle of ligand transfer from an ATP-binding cassette transporter LolCDE to a molecular chaperone LolA (2008)
  • Author(s): Tokuda, H.
  • Organizer: FEBS Special Meeting "ATP-binding cassette proteins : from multidrug resistance to genetic diseases"
  • Place of Presentation: Innsbruck, Austria
• [Presentation] Sorting of lipoproteins to the outer membrane of gram-negative bacteria. (2007)
  • Author(s): 福岡国際会議場, 福岡
  • Organizer: 007 ASBMB Annual Meeting
  • Place of Presentation: Washington DC Convention Center(USA)
  • Year and Date: 2007-04-29
• [Presentation] Sorting of lipoproteins to the outer membrane of Gram-negative bacteria (2007)
  • Author(s): Tokuda, H.
  • Organizer: The 2007 ASBMB meeting
  • Place of Presentation: Washington DC, USA
• [Book] Sorting of bacterial lipoproteins to the outer membrane by the Lolsystem. In "Methods in Molecular Biology ; Protein Secretion"(A. Economou (ed)) (2010)
  • Author(s): Narita, S., Tokuda, H.
  • Publisher: Humana press
• [Book] Structure, function and transport of lipoproteins in Escherichia coli. In "The Periplasm"(M. Ehrmann (ed)) (2007)
  • Author(s): Tokuda, H., Matsuyama, S., Tanaka-Masuda, K.
  • Publisher: ASM press
• [Book] The function of the ABC transporter LolCDE in protein transport to the outer membrane of E. coli. (2007)
  • Author(s): Narita, S., Tokuda, H.
  • Publisher: The Enzymes
• [Book] Biogenesis and membrane targeting of lipoproteins. In "E. coli and Salmonella"(Tracy Palmer (ed))
  • Author(s): Narita, S., Tokuda, H.
  • Publisher: ASM Press
• [Patent(Industrial Property Rights)] 大腸菌膜タンパク質挿入因子 (2010)
  • Inventor(s): 西山賢一, 徳田元, 植田卓也, 金森崇, 楠本正一, 前田将秀, 島本啓子
  • Industrial Property Rights Holder: サントリーホールディングス株式会社
  • Industrial Property Number: 2010-048520
  • Filing Date: 2010-03-05