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Structural basis for multidrug tripartite efflux pump through both the inner and outer membranes in Pseudomonas aeruginosa.

Research Project

Project/Area Number 25291014
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypePartial Multi-year Fund
Section一般
Research Field Structural biochemistry
Research InstitutionOsaka University

Principal Investigator

Yamashita Eiki  大阪大学, たんぱく質研究所, 助教 (00294132)

Co-Investigator(Kenkyū-buntansha) NAKAGAWA ATSUSHI  大阪大学, 蛋白質研究所, 教授 (20188890)
Research Collaborator YONEHARA RYO  大阪大学, 蛋白質研究所, 研究員
IKEDA ETSUKO  大阪大学, 蛋白質研究所, 博士研究員
堤 研太  大阪大学, 蛋白質研究所, 博士前期課程
HIGA YUKI  大阪大学, 蛋白質研究所, 博士前期課程
SUMITA KAZUMA  大阪大学, 蛋白質研究所, 博士前期課程
Project Period (FY) 2013-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥18,330,000 (Direct Cost: ¥14,100,000、Indirect Cost: ¥4,230,000)
Fiscal Year 2015: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2014: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2013: ¥10,920,000 (Direct Cost: ¥8,400,000、Indirect Cost: ¥2,520,000)
KeywordsX線結晶構造解析 / 膜タンパク質複合体の物質輸送 / 外膜チャンネルの構造
Outline of Final Research Achievements

The multidrug resistance of Pseudomonas aeruginosa is mainly attributable to high expression of the tripartite efflux pumps that transport antibiotics from within to the external environment. The tripartite efflux pumps, each of which is composed of an inner membrane transporter, a periplasmic adaptor protein and an outer membrane factor, are membrane-protein super-complexes spanning both the inner and outer membranes. In this study, we determined the stoichiometry of the tripartite efflux pump by size exclusion chromatography and gel electrophoresis and the structure of it by negative stain electron microscope. The structures of two major outer membrane factors from multidrug-resistance Pseudomonas aeruginosa were determined by X-ray crystallographic analysis.

Report

(4 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Annual Research Report
  • 2013 Annual Research Report
  • Research Products

    (20 results)

All 2016 2015 2014 2013 Other

All Journal Article (6 results) (of which Peer Reviewed: 6 results,  Acknowledgement Compliant: 1 results) Presentation (14 results) (of which Int'l Joint Research: 4 results,  Invited: 8 results)

  • [Journal Article] Crystal structures of OprN and OprJ, outer membrane factors of multidrug tripartite efflux pumps of Pseudomonas aeruginosa.2016

    • Author(s)
      Yonehara R, Yamashita E, Nakagawa A.
    • Journal Title

      Proteins.

      Volume: e-pub Issue: 6 Pages: 759-769

    • DOI

      10.1002/prot.25022

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Traffic within the Cytochrome b6f Lipoprotein Complex: Gating of the Quinone Portal.2014

    • Author(s)
      Hasan S. S., Proctor E. A., Yamashita E., Dokholyan N. V., Cramer W. A.
    • Journal Title

      Biophys J.

      Volume: 107 Issue: 7 Pages: 1620-628

    • DOI

      10.1016/j.bpj.2014.08.003

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Lipid-induced conformational changes within the cytochrome b6f complex of oxygenic photosynthesis.2013

    • Author(s)
      Hasan S. S., Stofleth J. T., Yamashita E., Cramer W. A.
    • Journal Title

      Biochemistry.

      Volume: 52 Issue: 15 Pages: 2649-2654

    • DOI

      10.1021/bi301638h

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A new protein complex promoting the assembly of Rad51 filaments.2013

    • Author(s)
      Sasanuma H., Tawaramoto M. S., Lao J. P., Hosaka H., Sanda E., Suzuki M., Yamashita E., Hunter N., Shinohara M., Nakagawa A., Shinohara A.
    • Journal Title

      Nat. Commun.

      Volume: 4 Issue: 1 Pages: 1676-1676

    • DOI

      10.1038/ncomms2678

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Crystallization and preliminary X-ray diffraction analysis of human importin β-Snail zinc finger domain complex.2013

    • Author(s)
      Choi S., Song J., Son S.Y., Park I.Y., Yamashita E., Lee S.J..
    • Journal Title

      Acta Cryst. F

      Volume: 69 Issue: 6 Pages: 1049-1051

    • DOI

      10.1042/bst20120211

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Crystal structure of the C-terminal domain of Mu phage central spike and functions of bound calcium ion.2013

    • Author(s)
      Harada K
    • Journal Title

      Biochim Biophys Acta.,

      Volume: 1834 Issue: 1 Pages: 284-291

    • DOI

      10.1016/j.bbapap.2012.08.015

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Presentation] Crystallography using Synchrotron Radiation2015

    • Author(s)
      Atsushi Nakagawa
    • Organizer
      Joint Symposium between the Institute for Protein Research (Osaka University) and the Research School of Chemistry (Australian National University) "Protein Structure and Function”
    • Place of Presentation
      Canberra(Australia)
    • Year and Date
      2015-11-14
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 緑膿菌由来異物排出トランスポーターMuxBのX 線結晶構造解析2015

    • Author(s)
      比嘉悠貴、米原 涼、 中川敦史、 山下栄樹
    • Organizer
      日本結晶学会年会
    • Place of Presentation
      大阪府立大学中百舌鳥キャンパス(大阪府堺市)
    • Year and Date
      2015-10-17
    • Related Report
      2015 Annual Research Report
  • [Presentation] Synchrotron radiation beam line of the Institute for Protein Research2015

    • Author(s)
      Eiki Yamashita
    • Organizer
      International Symposium on Establishment of Structural Biology Network in the Asian Region
    • Place of Presentation
      Hsinchu(Taiwan)
    • Year and Date
      2015-09-30
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 核外輸送に置ける Exportin-5 の構造基盤/The structural basis on nuclear export of Exportin-5.2015

    • Author(s)
      山澤龍治、小林武、慈幸千真理、月原冨武、中川敦史、山下栄樹
    • Organizer
      第15回 日本蛋白質科学会年会
    • Place of Presentation
      あわぎんホール(徳島県徳島市)
    • Year and Date
      2015-06-24
    • Related Report
      2015 Annual Research Report
  • [Presentation] Structure Biology Using Synchrotron Radiation2015

    • Author(s)
      Atsushi Nakagawa
    • Organizer
      Molecular and Cellular Life Sciences: Infectous Diseases, Biochemistry & Structural Biology (MCLS2015)
    • Place of Presentation
      Surabaya(Indonesia)
    • Year and Date
      2015-05-07
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Structure Determination of Biological Maromolecules by X-ray2015

    • Author(s)
      Atsushi Nakagawa
    • Organizer
      The First Trilateral Workshop for Frontier Protein Studies
    • Place of Presentation
      Peking(China)
    • Year and Date
      2015-04-23
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] SPring-8生体超分子複合体構造解析ビームライン(大阪大学蛋白質研究所)BL44XUの現状2015

    • Author(s)
      山下栄樹, 東浦彰史, 吉村政人, 長谷川和也, 熊坂崇,古川行人,大端通,上野剛,山本雅貴,吉川信也,月原冨武,中川敦史
    • Organizer
      第28回日本放射光学会年会放射光科学合同シンポジウム
    • Place of Presentation
      立命館大学びわこ・くさつキャンパス
    • Year and Date
      2015-01-10 – 2015-01-12
    • Related Report
      2014 Annual Research Report
  • [Presentation] 緑膿菌由来異物排出蛋白質のX線結晶構造解析2015

    • Author(s)
      住田一真,米原涼,中江太治,中川敦史,山下栄樹
    • Organizer
      第28回日本放射光学会年会放射光科学合同シンポジウム
    • Place of Presentation
      立命館大学びわこ・くさつキャンパス
    • Year and Date
      2015-01-10 – 2015-01-12
    • Related Report
      2014 Annual Research Report
  • [Presentation] IPR Beamline for Macromolecular Assemblies at SPring-8 (BL44XU)2014

    • Author(s)
      Eiki Yamashita, Akifumi Higashiura, Masato Yoshimura, Kazuya Hasegawa, Yukito Furukawa, Toru Ohata, Takashi Kumasaka, Go Ueno, Masaki Yamamoto, Atsushi Nakagawa
    • Organizer
      23rd Congress and General Assembly of the International Union of Crystallography
    • Place of Presentation
      Montreal
    • Year and Date
      2014-08-05 – 2014-08-12
    • Related Report
      2014 Annual Research Report
  • [Presentation] SPring-8生体超分子複合体構造解析ビームライン(大阪大学蛋白質研究所)BL44XUの現状2014

    • Author(s)
      山下栄樹
    • Organizer
      第27回日本放射光学会年会放射光科学合同シンポジウム
    • Place of Presentation
      広島国際会議場(広島)
    • Related Report
      2013 Annual Research Report
  • [Presentation] 構造生物学と放射光2013

    • Author(s)
      中川敦史
    • Organizer
      第13回日本蛋白質科学会年会
    • Place of Presentation
      とりぎん文化会館(鳥取)
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] Coherent Diffraction Imaging for Spherical Biological Particles2013

    • Author(s)
      Atsushi Nakagawa
    • Organizer
      The 28th European Crystallography Meeting
    • Place of Presentation
      Warick University, Coventry, UK
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] Coherent Diffraction Imaging for Structure Determination for Spherical Biological Particles2013

    • Author(s)
      Atsushi Nakagawa, Akifumi Higashiura, Kenji Iwasaki, Marina Murakami, Eiki Yamashita, Kazuki Takeda, Yu Hirano, Yuya Hanazono, Kiyofumi Takaba, Masahito Hibi, Yukiko Tomisaki, Kunio Miki
    • Organizer
      The 12th Conference of the Asian Crystallographic Association
    • Place of Presentation
      Hong Kong University of Science and Technology, Hong Kong
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] Structure determination of biological macromolecules using synchrotron radiation and X-ray free electron laser

    • Author(s)
      Atsushi Nakagawa
    • Organizer
      International Symposium on New Drug Discovery and Development
    • Place of Presentation
      Chungbuk University, Cheongju, Korea
    • Related Report
      2013 Annual Research Report
    • Invited

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Published: 2013-05-21   Modified: 2019-07-29  

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