Biogenesis of Escherichia coli cell envlope.

2011 Fiscal Year Final Research Report

• Project Area: Protein community: organization and maintenance of protein functions
• Project/Area Number: 19058003
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Morioka College (2010-2011); The University of Tokyo (2007-2009)
• Principal Investigator: TOKUDA Hajime 盛岡大学, 栄養科学部, 教授 (40125943)
• Co-Investigator(Kenkyū-buntansha): MATSUYAMA Shin-ichi (50183108) ; NISIYAMA Kennichi (80291334) ; NARITA Shin-ichiro (30338751) ; TAO Kazuyuki (00211996) ; TAKEDA Kazuki (30332290)
• Project Period (FY): 2007 – 2011
• Keywords: 外膜 / 細胞質膜 / リポ蛋白質 / βバレル型蛋白質 / Lol 因子 / リポ多糖

Research Abstract

Gram-negative bacteria generally possess outer membranes, which are essential for bacterial growth. In Escherichia coli, lipoproteins play critical roles in the biogenesis of outer membranes. Most E. coli lipoproteins are localized in the outer membrane. This is mediated by the Lol system comprising five Lol proteins in a manner dependent on lipoprotein sorting signals. The mechanisms underlying the biogenesis of E. coli outer membranes are examined by analyzing the outer membrane sorting of lipoproteins.

Research Products

• [Journal Article] Multiple SecA molecules drive protein translocation across a single translocon with SecG inversion. (2012)
  • Author(s): Morita, K., Tokuda, H., and Nishiyama, K.
  • Journal Title: J. Biol. Chem. Volume: 287 Pages: 455-464
  • Peer Reviewed
• [Journal Article] Defective lipoprotein sorting induces lolA expression through the Rcs stress response phosphorelay system. (2012)
  • Author(s): Tao, K., Narita, S., and Tokuda, H.
  • Journal Title: J. Bacteriol. Volume: (in press)
  • DOI: doi:10.1128/JB.00553-12
  • Peer Reviewed
• [Journal Article] Lipoprotein Sorting in Bacteria. (2011)
  • Author(s): Okuda, S., and Tokuda, H.
  • Journal Title: Annu. Rev. Microbiol. Volume: 65 Pages: 239-259
  • DOI: doi:10.1146/annurev-micro-090110-102859
  • Peer Reviewed
• [Journal Article] Overexpression of LolCDE allows deletion of the Escherichia coli gene encoding apolipoprotein N-acyltransferase. (2011)
  • Author(s): Narita, S., and Tokuda, H.
  • Journal Title: J. Bacteriol. Volume: 193 Pages: 4832-4840
  • DOI: doi:10.1128/JB.05013-11
  • Peer Reviewed
• [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., and Tokuda, H.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 394 Pages: 733-736
  • DOI: doi:10.1016/j.bbrc.2010.03.061
  • Peer Reviewed
• [Journal Article] Application of an inducible system to engineer unmarked conditional mutants of essential genes of Pseudomonas aeruginosa. (2010)
  • Author(s): Morita, Y., Narita, S., Tomida, J., Tokuda, H., and Kawamura, Y.
  • Journal Title: J. Microbiol. Methods Volume: 82 Pages: 205-213
  • DOI: doi:10.1016/j.mimet.2010.06.001
  • Peer Reviewed
• [Journal Article] A periplasmic LolA derivative with a lethal disulfide bond activates the Cpx stress response system. (2010)
  • Author(s): Tao, K., Watanabe, S., Narita, S., and Tokuda, H.
  • Journal Title: J. Bacteriol. Volume: 192 Pages: 5657-5662
  • DOI: doi:10.1128/JB.00821-10
  • Peer Reviewed
• [Journal Article] Preparation of a highly translocation-competent proOmpA/SecB complex. (2010)
  • Author(s): Nishiyama, K., and Tokuda, H.
  • Journal Title: Protein Sci. Volume: 19 Pages: 2402-2408
  • DOI: DOI:10.1002/pro.520
  • Peer Reviewed
• [Journal Article] Novel mutations of the LolCDE complex causing outer membrane localization of lipoproteins despite their inner membrane retention signals. (2010)
  • Author(s): Sakamoto, C., Satou, R., Tokuda, H., and Narita, S.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 401 Pages: 586-591
  • DOI: doi:10.1016/j.bbrc.2010.09.106
  • Peer Reviewed
• [Journal Article] Biogenesis of outer membranes in Gram-negative bacteria. (2009)
  • Author(s): Tokuda, H.
  • Journal Title: Biosci. Biotechnol. Biochem. Volume: 73 Pages: 465-473
  • DOI: DOI:10.1007/978-1-60327-412-8_7
  • Peer Reviewed
• [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., and Tokuda, H.
  • Journal Title: Proc. Natl. Acad. Sci., USA Volume: 106 Pages: 5877-5882
  • DOI: doi_10.1073_pnas.0900896106
  • 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., Mukaiyama, K., Okuda, S., Narita, S., and Tokuda, H.
  • Journal Title: FEBS J. Volume: 276 Pages: 4496-4504
  • DOI: doi:10.1111/j.1742-4658.2009.07156.x
  • Peer Reviewed
• [Journal Article] Biochemical characterization of an ABC transporter LptBFGC complex required for the outer membrane sorting of lipopolysaccharides. (2009)
  • Author(s): Narita, S., and Tokuda, H.
  • Journal Title: FEBS Lett. Volume: 583 Pages: 2160-2164
  • DOI: doi:10.1016/j.febslet.2005.10.038
  • Peer Reviewed
• [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., and Shimada, I.
  • Journal Title: J. Biol. Chem. Volume: 284 Pages: 24634-24643
  • DOI: DOI:10.1074/jbc.M109.001149
  • Peer Reviewed
• [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., and Narita, S.
  • Journal Title: Biosci. Biotechnol. Biochem. Volume: 73 Pages: 2310-2316
  • DOI: DOI:10.1074/jbc.M109.001149
  • Peer Reviewed
• [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., and Tokuda, H.
  • Journal Title: FEBS Lett. Volume: 583 Pages: 2987-2990
  • DOI: doi:10.1016/j.febslet.2009.08.032
  • Peer Reviewed
• [Journal Article] Development of a functional in vitro integration system for an integral membrane protein, SecG. (2009)
  • Author(s): Nishiyama, K., and Tokuda, H.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 390 Pages: 920-924
  • DOI: doi:10.1016/j.bbrc.2009.10.078
  • 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., and Tokuda, H.
  • Journal Title: J. Biol. Chem. Volume: 283 Pages: 8538-8544
  • DOI: DOI:10.1074/jbc.M800026200
  • Peer Reviewed
• [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., and Tokuda, H.
  • Journal Title: FEBS Lett. Volume: 582 Pages: 2247-2251
  • DOI: doi:10.1016/j.febslet.2008.05.022
  • Peer Reviewed
• [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., and Nishiyama, K.
  • Journal Title: J. Biol. Chem. Volume: 283 Pages: 24489-24496
  • DOI: DOI:10.1074/jbc.M80812200
  • Peer Reviewed
• [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., and Tokuda, H.
  • Journal Title: J. Biol. Chem. Volume: 283 Pages: 25414-25420
  • DOI: DOI:10.1074/jbc.M804736200
  • 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., Oguchi, Y., Takeda, K., Miki, K., and Tokuda, H.
  • Journal Title: J. Biol. Chem. Volume: 283 Pages: 25421-25427
  • DOI: DOI:10.1074/jbc.M804737200
  • 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., and Tokuda, H.
  • Journal Title: Microbiol. Immunol. Volume: 51 Pages: 263-270
  • URL: http://onlinelibrary.wiley.com/journal/10.1111/(ISSN)1348-0421
  • Peer Reviewed
• [Journal Article] Amino acids at positions 3 and 4 determine the membrane specificity of Pseudomonas aeruginsoa lipoproteins. (2007)
  • Author(s): Narita, S., and Tokuda, H.
  • Journal Title: J. Biol. Chem. Volume: 282 Pages: 13372-13378
  • DOI: DOI:10.1074/jbc.M611839200
  • Peer Reviewed
• [Journal Article] Characterization of the Pseudomonas aeruginosa Lol system as a lipoprotein sorting mechanism. (2007)
  • Author(s): Tanaka, S., Narita, S., and Tokuda, H.
  • Journal Title: J. Biol. Chem. Volume: 282 Pages: 13379-13384
  • DOI: DOI:10.1074/jbc.M611840200
  • Peer Reviewed
• [Journal Article] Diverse effects of phospholipids on lipoprotein sorting and ATP hydrolysis by the ABC transporter LolCDE complex. (2007)
  • Author(s): Miyamoto, S., and Tokuda, H.
  • Journal Title: Biochim. Biophys. Acta-Biomembranes Volume: 1768 Pages: 1848-1854
  • DOI: doi:10.1016/j.bbamem.2007.04.005
  • Peer Reviewed
• [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., and Tokuda, H.
  • Journal Title: FEBS J. Volume: 274 Pages: 3034-3043
  • DOI: doi:10.1111/j.1742-4658.2007.05832.x
  • Peer Reviewed
• [Journal Article] Structural studies of the Cpx pathway activator NlpE on the outer membrane of Escherichia coli. (2007)
  • Author(s): Hirano, Y., Hossain, Md. M., Takeda, K., Tokuda, H., and Miki, K.
  • Journal Title: Structure Volume: 15 Pages: 963-976
  • DOI: DOI:10.1016/j.str.2007.06.014
  • Peer Reviewed
• [Journal Article] Topology inversion of SecG is essential for the cytosolic SecA-dependent stimulation of protein translocation. (2007)
  • Author(s): Sugai, R., Takemae, K., Tokuda, H., and Nishiyama, K.
  • Journal Title: J. Biol. Chem. Volume: 282 Pages: 29540-29548
  • DOI: DOI:10.1074/jbc.M704716200
  • Peer Reviewed
• [Journal Article] Backbone resonance assignment for the outer membrane lipoprotein receptor LolB from Escherichia coli. (2007)
  • Author(s): Nakada, S., Sakakura, M., Takahashi, H., Tokuda, H., and Shimada, I.
  • Journal Title: Biomol. NMR Assign. Volume: 1 Pages: 121-123
  • DOI: DOI:10.1074/jbc.M109.001149
  • 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-dependent manner. (2007)
  • Author(s): Watanabe, S., Oguchi, Y., Yokota, N., and Tokuda, H.
  • Journal Title: Protein Sci. Volume: 16 Pages: 2741-2749
  • DOI: doi:10.1110/ps.073101307
  • Peer Reviewed
• [Presentation] How does the LolCDE complex recognize and release bacterial lipoproteins from the cytoplasmic membrane? (2011)
  • Author(s): Tokuda, H.
  • Organizer: The JSPS/iCeMS International Symposium“ ABC2011 in Kyoto-ABC Proteins/Membrane Meso-domains/ES-iPS cells”
  • Place of Presentation: Kyoto University, Japan
  • Year and Date: 20111116-17
• [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: 20100227-0305
• [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: 20080301-08
• [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: 20070428-0502
• [Book] Sorting of bacterial lipoproteins to the outer membrane by the Lol system. In “Methods in Molecular Biology; Protein Secretion”, A. Economou (ed), DOI:10.1007/978-1-60327-412-8_7, (2010)
  • Author(s): Narita, S., and Tokuda, H.
  • Total Pages: 117-129
  • Publisher: Humana press
• [Book] The function of the ABC transporter LolCDE in protein transport to the outer membrane of E. coli. In The Enzymes, vol. 25, DOI:10.1016/S1874-6047(07)25007-3 (2007)
  • Author(s): Narita, S., and Tokuda, H.
  • Total Pages: 149-172
• [Book] ,Structure, function and transport of lipoproteins in Escherichia coli. In “The Periplasm”, M. Ehrmann (ed) (2007)
  • Author(s): Tokuda, H., Matsuyama, S., and Tanaka-Masuda, K.
  • Total Pages: 67-79
  • Publisher: ASM press
• [Book] Bacterial lipoproteins; biogenesis, trafficking and function. In press Chapter 4 in Bacterial Membranes: Structural and Molecular Biology Han Remaut and Remi Fronzes, eds,
  • Author(s): Tokuda, H., Sander, P., Lee, B. L., Okuda, S., Grau, T., Tschumi, A., Brulle, J. K., Kurokawa, K., and Nakayama, H.
  • Publisher: Horizon Scientific Press
• [Book] Biogenesis and membrane targeting of lipoproteins. Chapter 4.3.7. In “Escherichia coli and Salmonella: cellular and molecular biology.” Bock, R. Curtiss III, J. B. Kaper, P. D. Karp, F. C. Neidhardt, T. Nystrom, J. M. Slauch, and C. L. Squires, and D. Ussery (ed), doi:10.1128/ecosal.4.3.7 (online journal).
  • Author(s): Narita, S., and Tokuda, H.
  • Publisher: ASM Press. Washington, DC.
• [Remarks]
  • URL: http://www.iam.u-tokyo.ac.jp/sec/index-3.html
• [Remarks]
  • URL: http://www.morioka-u.ac.jp/UV_ns/tokuda_lab/