Clarification of the physiological role of diacylglycerol in E. coli

• Project/Area Number: 21570110
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Iwate University
• Principal Investigator: NISHIYAMA Ken-ichi 岩手大学, 農学部, 教授 (80291334)
• Project Period (FY): 2009 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2012: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2010: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2009: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: MPIase / SecG / ジアシルグリセロール / タンパク質膜挿入 / 再構成 / 大腸菌 / 自発的膜挿入 / 低温感受性 / 機能的膜タンパク質合成

Research Abstract

To clarify the physiological role of diacylglycerol in E. coli, ～10 genes, which may be involved in diacylglycerol biosynthesis, were listed up, and then constructed multiply knock-out strains. Resultantly, an eightfold knock-out strain could be constructed, of which the growth was cold-sensitive and the diacylglycerol content was greatly reduced. However, the depletion was not complete. On the other hand, we determined the structure and analyzed the function of glycolipid MPIase involved in protein integration into membranes.

Research Products

• [Journal Article] Glycolipozyme MPIase is essential for topology inversion of SecG during preprotein translocation (2012)
  • Author(s): Moser, M., Nagamori, S., Huber, M., Tokuda, H., Nishiyama, K.
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: (in press) Issue: 24 Pages: 9734-9739
  • DOI: 10.1073/pnas.1303160110
  • Peer Reviewed
• [Journal Article] MPIase is a glycolipozyme essential for membrane protein integration (2012)
  • Author(s): Nishiyama, K., Maeda,M., Yanagisawa,K., Nagase, R., Komura, H., Iwashita, T., Yamagaki, T., Kusumoto, S., Tokuda, H., Shimamoto, K.
  • Journal Title: Nat. Commun Volume: 3 Issue: 1 Pages: 1-10
  • DOI: 10.1038/ncomms2267
  • NAID: 120007179832
  • Peer Reviewed
• [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巻 Issue: 1 Pages: 455-464
  • DOI: 10.1074/jbc.m111.301754
  • Peer Reviewed
• [Journal Article] Multiple SecA molecules drive protein translocation across a single translocon with SecG inversion (2012)
  • Author(s): Nishiyama, K (Morita, K)
  • Journal Title: J Biol Chem Volume: 287 Pages: 455-464
  • 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 Issue: 12 Pages: 2402-2408
  • DOI: 10.1002/pro.520
  • 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 Issue: 3 Pages: 733-736
  • DOI: 10.1016/j.bbrc.2010.03.061
  • NAID: 120002439245
  • Peer Reviewed
• [Journal Article] A novel complete reconstitution system for membrane integration of the simplest membrane protein (2010)
  • Author(s): 西山賢一
  • Journal Title: Biochem. Biophys. Res.Commun. Volume: 394 Pages: 733-736
  • NAID: 120002439245
  • Peer Reviewed
• [Journal Article] Preparation of a highly translocation-competent proOmpA/SecB complex (2010)
  • Author(s): 西山賢一
  • Journal Title: Protein Science Volume: 19 Pages: 2402-2408
  • Peer Reviewed
• [Journal Article] A novel complete reconstitution system for membrane integration of the simplest membrane protein (2010)
  • Author(s): 西山賢一
  • Journal Title: Biochem.Biophys.Res.Commun. 394(印刷中)
  • NAID: 120002439245
  • 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 Issue: 3 Pages: 920-924
  • DOI: 10.1016/j.bbrc.2009.10.078
  • Peer Reviewed
• [Journal Article] Development of a functional in vitro integration system for an integral membrane protein, SecG (2009)
  • Author(s): 西山賢一
  • Journal Title: Biochem.Biophys.Res.Commun. 390 Pages: 920-924
  • Peer Reviewed
• [Presentation] 大腸菌のタンパク質膜挿入に必須の糖脂質酵素(Glycolipozyme) MPIaseの構造と機能 (2013)
  • Author(s): 西山賢一
  • Organizer: 日本農芸化学会2013年度大会 シンポジウム「グラム陰性細菌細胞表層構造の形成機構とバイオテクノロジー」
  • Place of Presentation: 東北大学川内北キャンパス(宮城県)
  • Year and Date: 2013-03-27
• [Presentation] 大腸菌のタンパク質膜挿入に必須の糖脂質酵素（Glycolipozyme） MPIaseの構造と機能 (2013)
  • Author(s): 西山賢一
  • Organizer: 日本農芸化学会2013年度大会 シンポジウム「グラム陰性細菌細胞表層構造の形成機構とバイオテクノロジー」
  • Place of Presentation: 宮城県仙台市、東北大学川内北キャンパス
• [Presentation] 大腸菌のタンパク質膜挿入に必須の糖脂質酵素(Glycolipozyme) MPIaseの構造と機能 (2012)
  • Author(s): 西山賢一
  • Organizer: 第7回無細胞生命科学研究会
  • Place of Presentation: 愛媛大学南加記念ホール(愛媛県)
  • Year and Date: 2012-11-17
• [Presentation] タンパク質膜挿入に必須の糖脂質酵素MPIaseの構造と機能 (2012)
  • Author(s): 西山賢一
  • Organizer: 第9回21世紀大腸菌研究会
  • Place of Presentation: 長浜ロイヤルホテル(滋賀県)
  • Year and Date: 2012-06-22
• [Presentation] MPIase as a glycolipozyme essential for membrane protein integration (2012)
  • Author(s): Ken-ichi Nishiyama
  • Organizer: Gordon Research Conference on Bacterial Cell Surface
  • Place of Presentation: Mount Snow Resort, West Dover, VT, USA
• [Presentation] タンパク質膜挿入に必須の糖脂質MPIaseとその標的タンパク質との相互作用解析 (2012)
  • Author(s): Michael Moser（西山賢一）
  • Organizer: 新学術領域研究「天然物ケミカルバイオロジー～分子標的と活性制御～」第2回公開シンポジウム
  • Place of Presentation: 東京都、東京大学弥生講堂
• [Presentation] タンパク質膜挿入に必須の糖脂質酵素MPIaseの構造と機能 (2012)
  • Author(s): 西山賢一
  • Organizer: 第9回21世紀大腸菌研究会
  • Place of Presentation: 滋賀県長浜市、長浜ロイヤルホテル
• [Presentation] MPIase as a glycolipozyme essential for membrane protein integration (2012)
  • Author(s): Ken-ichi Nishiyama
  • Organizer: Gordon Research Conference on Bacterial Cell Surface
  • Place of Presentation: Mount Snow Resort, Wset Dover, VT, USA
• [Presentation] MPIase, a novel glycolipozyme, is essential for topology inversion of SecG (2012)
  • Author(s): Michael Moser (Nishiyama, K)
  • Organizer: The 1st International Symposium on Chemical Biology of Natural Products: Target ID and Regulation of Bioactivity
  • Place of Presentation: 京都市、京都センチュリーホテル
• [Presentation] Structure and function of novel glycolipid characterized as membrane protein integrase (2012)
  • Author(s): Keiko Shimamoto (Nishiyama, K)
  • Organizer: The 1st International Symposium on Chemical Biology of Natural Products: Target ID and Regulation of Bioactivity
  • Place of Presentation: 京都市、京都センチュリーホテル
• [Presentation] タンパク質膜挿入に必須の糖脂質酵素（Glycolipozyme）MPIase (2012)
  • Author(s): 西山賢一
  • Organizer: 第7回 無細胞生命科学研究会プログラム
  • Place of Presentation: 愛媛県松山市、愛媛大学南加記念ホール
• [Presentation] タンパク質膜挿入・膜透過に関与する糖脂質MPIase の構造と機能 (2011)
  • Author(s): 西山賢一
  • Organizer: 第6回無細胞生命科学研究会
  • Place of Presentation: 兵庫県立大学(兵庫県)
  • Year and Date: 2011-11-16
• [Presentation] タンパク質膜挿入に関与する新規複合糖脂質MPIaseの構造と機能 (2011)
  • Author(s): 西山賢一
  • Organizer: 第84回生化学会大会
  • Place of Presentation: 京都国際会館(京都)
  • Year and Date: 2011-09-24
• [Presentation] MPIaseによるSecYEGの構造変化とSecGの膜内配向性反転 (2010)
  • Author(s): 西山賢一
  • Organizer: 第5回無細胞生命科学研究会
  • Place of Presentation: 岡山大学
  • Year and Date: 2010-09-29
• [Presentation] MPIase, a novel glycolipid involved in membrane protein integration and preprotein translocation (2010)
  • Author(s): Ken-ichi Nishiyama
  • Organizer: The 3rd International Symposium on Protein Community
  • Place of Presentation: ホテル日航奈良(奈良)
  • Year and Date: 2010-09-14
• [Presentation] MPIase, a novel glycolipid involved in membrane protein integration and preprotein translocation (2010)
  • Author(s): 西山賢一
  • Organizer: The 3rd International Symposiun on protein Community
  • Place of Presentation: 奈良、ホテル日航 招待講演
  • Year and Date: 2010-09-14
• [Presentation] A novel complete reconstitution system for membrane integration of the simplest membrane protein (2010)
  • Author(s): Ken-ichi Nishiyama
  • Organizer: Gordon Research Conference on Bacterial Cell Surfaces
  • Place of Presentation: New London, NH, USA
  • Year and Date: 2010-06-30
• [Presentation] 機能的膜タンパク質のin vitro合成と膜挿入 (2009)
  • Author(s): 西山賢一
  • Organizer: 第4回無細胞生命科学研究会
  • Place of Presentation: 下呂市民会館(岐阜)
  • Year and Date: 2009-11-16
• [Presentation] 大腸菌におけるタンパク質膜挿入機構 (2009)
  • Author(s): 西山賢一
  • Organizer: 第6回21世紀大腸菌研究会
  • Place of Presentation: KKRホテル熱海(静岡)
  • Year and Date: 2009-06-12
• [Presentation] The oligomeric structure of the E.coil SecYEG translocon is influenced by MPIase, a novel glycolipid
  • Author(s): 西山賢一
  • Organizer: 第84回生化学会大会
  • Place of Presentation: 京都国際会館
• [Presentation] タンパク質膜挿入に関与する新規複合糖脂質MPIaseの構造と機能
  • Author(s): 西山賢一
  • Organizer: 第84回生化学会大会
  • Place of Presentation: 京都国際会館
• [Presentation] Construction of SecYEG translocon by cell-free protein synthesis system
  • Author(s): 西山賢一
  • Organizer: 第84回生化学会大会
  • Place of Presentation: 京都国際会館
• [Presentation] 膜タンパク質挿入に関わる新規糖脂質MPIaseの機能と構造
  • Author(s): 西山賢一
  • Organizer: 第53回天然有機化合物討論会
  • Place of Presentation: 大阪国際センター(招待講演)
• [Presentation] Cell-Free Construction of Synthetic Biomembrane
  • Author(s): 西山賢一
  • Organizer: Annual Meeting of Cell Synthesis Research
  • Place of Presentation: 千里ライフサイエンスセンター
• [Presentation] タンパク質膜挿入・膜透過に関与する糖脂質MPIaseの構造と機能
  • Author(s): 西山賢一
  • Organizer: 第6回無細胞生命科学研究会
  • Place of Presentation: 兵庫県立大学
• [Presentation] 抗MPIase抗体を用いた糖脂質MPIaseの構造と機能解析
  • Author(s): 西山賢一
  • Organizer: 第6回無細胞生命科学研究会
  • Place of Presentation: 兵庫県立大学
• [Presentation] 大腸菌膜タンパク質挿入に関わる新しい因子:機能と構造
  • Author(s): 西山賢一
  • Organizer: 第14回生命化学研究会
  • Place of Presentation: ラフォーレ南紀白浜(招待講演)
• [Book] 膜タンパク質の鍵は糖脂質にあり-すべての生体膜挿入に必須な因子を求めて-月刊化学 (2013)
  • Author(s): 西山賢一、島本啓子
  • Publisher: 化学同人
• [Remarks]
  • URL: http://news7a1.atm.iwate-u.ac.jp/~sec/
• [Remarks]
  • URL: http://news7al.atm.iwate-u.ac.jp/~sec/
• [Remarks]
  • URL: http://news7a1.atm.iwate-u.ac.jp/~sec/
• [Patent(Industrial Property Rights)] 大腸菌膜タンパク質挿入因子 (2011)
  • Inventor(s): 楠本正一、島本啓子、前田将秀、西山賢一、徳田元、上田卓也、金森崇
  • 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: 2011-03-05
• [Patent(Industrial Property Rights)] 大腸菌膜タンパク質挿入因子 (2010)
  • Inventor(s): 楠本正一, 島本啓子, 前田将秀, 西山賢一, 徳田元, 上田卓也, 金森崇
  • 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