Investigation of Chaperone Function of Phospholipids Containing Polyunsaturated Fatty Acids and Their Application to Overproduction of Membrane Proteins

• Project/Area Number: 22658028
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Applied microbiology
• Research Institution: Kyoto University
• Principal Investigator: KURIHARA Tatsuo 京都大学, 化学研究所, 教授 (70243087)
• Co-Investigator(Renkei-kenkyūsha): KAWAMOTO Jun 京都大学, 化学研究所, 助教 (90511238)
• Project Period (FY): 2010 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥3,560,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥360,000); Fiscal Year 2011: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2010: ¥2,000,000 (Direct Cost: ¥2,000,000)
• Keywords: 微生物 / 生体膜 / 高度不飽和脂肪酸 / エイコサペンタエン酸 / リン脂質 / 膜タンパク質 / フォールディング / ポーリン

Research Abstract

Shewanella livingstonensis Ac10 produces eicosapentaenoic acid-containing phospholipids (EPA-PLs) at low temperatures. We analyzed the role of EPA-PLs in the folding of Omp74, a major cold-inducible outer membrane protein of this bacterium. We found that the conformation of Omp74 in the mutant lacking EPA-PLs is different from that in the wild-type strain. In vitro experiments using liposomes showed that EPA-PLs function as a chemical chaperone in the membrane integration and folding of Omp74.

Research Products

• [Journal Article] Eicosapentaenoic acid facilitates the folding of an outer membrane protein of the psychrotrophic bacterium (2012)
  • Author(s): Xian-Zhu Dai, Jun Kawamoto, Satoshi B. Sato, Nobuyoshi Esaki, and Tatsuo Kurihara
  • Journal Title: Shewanella livingstonensis Ac10. Biochem. Biophys. Res. Commun. Volume: 425 Issue: 2 Pages: 363-7
  • DOI: 10.1016/j.bbrc.2012.07.097
  • Peer Reviewed
• [Journal Article] Occurrence of bacterial membrane microdomain at the cell division site enriched in phospholipids with polyunsaturated hydrocarbon chain. (2012)
  • Author(s): Sato, S., Kawamoto, J., Sato, S.B., Watanabe, B., Hiratake, J., Esaki, N., and Kurihara, T.
  • Journal Title: Journal of Biological Chemistry Volume: 287 Issue: 29 Pages: 24113-21
  • DOI: 10.1074/jbc.m111.318311
  • NAID: 120005302081
  • Peer Reviewed
• [Journal Article] dentification of cold-inducible inner membrane proteins of the psychrotrophic bacterium, Shewanella livingstonensis Ac10, by proteomic analysis. (2012)
  • Author(s): Park, J., Kawamoto, J., Esaki, N., and Kurihara, T.
  • Journal Title: Extremophiles Volume: 16 Issue: 2 Pages: 227-236
  • DOI: 10.1007/s00792-011-0422-z
  • Peer Reviewed
• [Journal Article] Characterization of 1-acyl-sn-glycerol-3-phosphate acyltransferase from a polyunsaturated fatty acid-producing bacterium (2012)
  • Author(s): Hyun-Nam Cho, Wataru Kasai, Jun Kawamoto, Nobuyoshi Esaki, and Tatsuo Kurihara
  • Journal Title: Shewanella livingstonensis Ac10. Trace Nutrients Research Volume: 29 Pages: 92-9
  • URL: http://www.jtnrs.com/sym29/19-No-P-14.pdf
  • Peer Reviewed
• [Journal Article] Functional analysis of an eicosapentaenoic acid biosynthesis protein Orf2 from a psychrotrophic bacterium (2012)
  • Author(s): Chunjie Gong, Jun Kawamoto, Nobuyoshi Esaki, and Tatsuo Kurihara
  • Journal Title: Functional analysis of an eicosapentaenoic acid biosynthesis protein Orf2 from a psychrotrophic bacterium Volume: 29 Pages: 84-91
  • URL: http://www.jtnrs.com/sym29/18-No-P-13.pdf
  • Peer Reviewed
• [Journal Article] Eicosapentaenoic acid plays a role in stabilizing dynamic membrane structure in the deep-sea piezophile Shewanella violacea (2012)
  • Author(s): Usui, Keiko, et al
  • Journal Title: Biochim.Biophys.Acta Volume: 1818 Pages: 574-83
  • Peer Reviewed
• [Journal Article] In vitro refolding of an OmpA homolog, a major cold-inducible outer membrane protein, from a psychrotrophic bacterium, Shewanella livingstonensis Ac10 (2011)
  • Author(s): Xian-Zhu Dai, Jun Kawamoto, NobuyoshiEsaki, and Tatsuo Kurihara
  • Journal Title: Journal of Japanese Society for Extremophiles Volume: 10 Pages: 90-6
  • Peer Reviewed
• [Journal Article] Favourable effects of eicosapentaenoic acid on the late step of the cell division in a piezophilic bacterium, Shewanella violacea DSS12, at high-hydrostatic pressures (2011)
  • Author(s): Kawamoto, J., Sato, T., Nakasone, K., Kato, C., Mihara, H., Esaki, N., and Kurihara, T
  • Journal Title: Environmental Microbiology Volume: 13 Issue: 8 Pages: 2293-2298
  • DOI: 10.1111/j.1462-2920.2011.02487.x
  • Peer Reviewed
• [Journal Article] In vitro refolding of an OmpA homolog, a major cold-inducible outer membrane protein, from a psychrotrophic bacterium, Shewanella livingstonensis Ac10 (2011)
  • Author(s): Dai, Xian-Zhu, et al
  • Journal Title: Journal of Japanese Society for Extremophiles Volume: 10 Pages: 90-96
  • Peer Reviewed
• [Journal Article] Favourable effects of eicosapentaenoic acid on the late step of the cell division in a piezophilic bacterium, Shewanella violaceg DSS12, at high-hydrostatic pressures (2011)
  • Author(s): Kawamoto, Jun, et al
  • Journal Title: Environ.Microbiol. Volume: 13 Pages: 2293-2298
  • Peer Reviewed
• [Presentation] 化学的アプローチによる高度不飽和脂肪酸含有リン脂質の生理機能解析 (2013)
  • Author(s): 栗原達夫
  • Organizer: 日本薬学会第133年会
  • Place of Presentation: 横浜市 パシフィコ横浜
  • Year and Date: 2013-03-29
• [Presentation] 細菌における高度不飽和脂肪酸含有リン脂質の生合成と機能 (2013)
  • Author(s): 栗原達夫
  • Organizer: 日本農芸化学会2013年度大会
  • Place of Presentation: 仙台市 東北大学
  • Year and Date: 2013-03-27
• [Presentation] 好冷性細菌の低温適応を担う分子基盤:細胞膜における高度不飽和脂肪酸の機能 (2012)
  • Author(s): 栗原達夫
  • Organizer: 新化学技術推進協会講演会
  • Place of Presentation: 京都市 京都テルサ
  • Year and Date: 2012-11-29
• [Presentation] Shewanella livingstonensis Ac10 におけるエイコサペンタエン酸の機能 (2012)
  • Author(s): 栗原達夫
  • Organizer: 特殊環境微生物セミナー
  • Place of Presentation: 特殊環境微生物セミナー
  • Year and Date: 2012-11-22
• [Presentation] Function of eicosapentaenoic acid-containing phospholipids in bacterial cell membrane (2012)
  • Author(s): 栗原達夫
  • Organizer: Hirschegg Summer School 2012
  • Place of Presentation: Marburger Haus,Hirschegg, Austria
  • Year and Date: 2012-09-06
• [Presentation] 生体膜における高度不飽和脂肪酸含有リン脂質の機能 (2012)
  • Author(s): 栗原達夫
  • Organizer: 京都大学学際融合教育研究推進センター生理化学研究ユニット第2回シンポジウム
  • Place of Presentation: 京都市 京都大学
  • Year and Date: 2012-06-03
• [Presentation] 細菌細胞膜における高度不飽和脂肪酸含有リン脂質の機能:局在化と膜タンパク質生合成における機能 (2012)
  • Author(s): 栗原達夫
  • Organizer: 日本農芸化学会2012年度大会
  • Place of Presentation: 京都市 京都女子大学
  • Year and Date: 2012-03-25
• [Presentation] 細菌細胞膜におけるリン脂質の局在化と生理機能 (2011)
  • Author(s): 栗原達夫
  • Organizer: 京都大学微生物科学寄附研究部門主催シンポジウム「微生物科学研究の現状と展望」
  • Place of Presentation: 京都市 芝蘭会館
  • Year and Date: 2011-06-23
• [Presentation] 細菌細胞膜におけるリン脂質の局在化と生理機能 (2011)
  • Author(s): 栗原達夫
  • Organizer: 京都大学微生物科学寄附研究部門主催シンポジウム
  • Place of Presentation: 京都市・芝蘭会館(招待講演)
  • Year and Date: 2011-06-23
• [Presentation] Function of phospholipids containing eicosapentaenoic acid in the bacterial cell membrane (2011)
  • Author(s): 栗原達夫
  • Organizer: Wageningen University and Research Centre/Kyoto University Joint Workshop
  • Place of Presentation: 京都市 京都大学
  • Year and Date: 2011-04-26
• [Presentation] Molecular probes to analyze the dynamics of eicosapentaenoic-acid-containing phospholipids and their interaction with membrane proteins (2011)
  • Author(s): 栗原達夫
  • Organizer: 第30回内藤コンファレンス
  • Place of Presentation: 札幌市 シャトレーゼ ガトーキングダムサッポロ
• [Presentation] Construction of protein expression system by using cold-adapted bacteria as the host (2010)
  • Author(s): 栗原達夫
  • Organizer: MicroPerm Workshop
  • Place of Presentation: Alfred Wegener Institute for Polar and Marine Research, Potsdam, Germany
  • Year and Date: 2010-11-08