Molecular mechanisms underlying the membrane sorting of bacterial lipoproteins

2009 Fiscal Year Final Research Report

• 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
• 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

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
• [Journal Article] Biogenesis of outer membranes in Gram-negative bacteria (2009)
  • Author(s): Tokuda, H.
  • Journal Title: Biosci.Biotechnol.Biochem. 73 Pages: 465-473
• [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
• [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] 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] 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
• [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
• [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
  • Year and Date: 20100227-20100305
• [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
  • Year and Date: 20080301-20080308
• [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
  • Year and Date: 20070428-20070502
• [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.
  • Total Pages: 117-129
  • 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.
  • Total Pages: 67-79
  • 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.
  • Total Pages: 149-172
  • 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: 特許特願2010-048520
  • Filing Date: 2010-03-05