Elucidation of membrane protein integration mechanism of glycolipid MPIase by using synthetic MPIase library.

• Project/Area Number: 17K13262
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Biomolecular chemistry
• Research Institution: Suntory Foundation for Life Sciences
• Principal Investigator: Fujikawa Kohki 公益財団法人サントリー生命科学財団, 生物有機科学研究所・構造生命科学研究部, 研究員 (50755874)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2019: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 糖脂質 / 膜タンパク質 / 膜挿入 / シャペロン / グリコシル化

Outline of Final Research Achievements

To elucidate the membran protein integration mechanism of glycolipid MPIase, which has the function of inserting membrane proteins of Escherichia coli into membranes, chemical synthesis of various types of analogs, including trisaccharyl pyrophospholipid (mini-MPIase-3), was carried out and it was used for structure-activity relationship studies. As a result, the membrane protein integration mechanism in which the sugar chain of MPIase captures the membrane protein and leads to the inner membrane by suppressing protein aggregation was demonstrated. In addition, the sugar chain of MPIase has chaperone-like activity, GlcNAc 6-OAc group, a carboxy group, and pyrophosphate are essential for the activity, and anchoring MPIase to the membrane contribute increasing the fluidity of the membrane were proved.

Academic Significance and Societal Importance of the Research Achievements

これまでに糖脂質が膜タンパク質の膜挿入に関与しているという報告はなく、糖脂質の酵素・シャペロン様の新機能を示すことができた。糖鎖のシャペロン様活性は、タンパク質の凝集抑制(可溶化)剤の開発につながり、タンパク質凝集が引き金になるアルツハイマー病などの疾患の解決にも応用が期待される。また、膜タンパク質を膜挿入する合成分子を初めて創出できた事から、今後、無細胞タンパク質合成系を用いた膜タンパク質の再構成効率を高める分子が多く開発され、再構成の難しい膜タンパク質研究が推進されると期待される。

Research Products

• [Journal Article] Novel Glycolipid Involved in Membrane Protein Integration: Structure and Mode of Action (2019)
  • Author(s): Fujikawa, K.; Nomura, K.; Nishiyama, K. I.; Shimamoto, K.
  • Journal Title: Journal of Synthetic Organic Chemistry, Japan Volume: 77 Issue: 11 Pages: 1096-1105
  • DOI: 10.5059/yukigoseikyokaishi.77.1096
  • NAID: 130007742536
  • ISSN: 0037-9980, 1883-6526
  • Year and Date: 2019-11-01
  • Peer Reviewed / Open Access
• [Journal Article] Enzyme-like ｇlycolipids MPIase involved in membrane protein integration in E. coli (2019)
  • Author(s): Kohki Fujikawa, Ken-ichi Nishiyama, Keiko Shimamoto
  • Journal Title: Trends in Glycoscience and Glycotechnology Volume: 印刷中
  • NAID: 130007751011
  • Peer Reviewed / Open Access
• [Journal Article] Glycan Derivatives That Possess Chaperone-Like Activity (2018)
  • Author(s): Kohki Fujikawa
  • Journal Title: Trends in Glycoscience and Glycotechnology Volume: 30 Issue: 177 Pages: E241-E243
  • DOI: 10.4052/tigg.1831.6E
  • NAID: 130007521404
  • ISSN: 0915-7352, 1883-2113
  • Year and Date: 2018-11-25
  • Peer Reviewed / Open Access
• [Journal Article] Syntheses and Activities of the Functional Structures of a Glycolipid Essential for Membrane Protein Integration (2018)
  • Author(s): Fujikawa Kohki、Suzuki Sonomi、Nagase Ryohei、Ikeda Shiori、Mori Shoko、Nomura Kaoru、Nishiyama Ken-ichi、Shimamoto Keiko
  • Journal Title: ACS Chemical Biology Volume: 13 Issue: 9 Pages: 2719-2727
  • DOI: 10.1021/acschembio.8b00654
  • Peer Reviewed / Open Access
• [Journal Article] グリコシル化反応における水素結合 (2017)
  • Author(s): 藤川紘樹
  • Journal Title: Trends in Glycoscience and Glycotechnology Volume: 29
  • NAID: 130006219285
  • Open Access
• [Presentation] 大腸菌膜タンパク質膜挿入に必須な糖脂質MPIase類縁体の合成 (2019)
  • Author(s): 藤川紘樹, 鈴木苑実, 池田汐里, 森祥子, 野村薫, 西山賢一, 島本啓子
  • Organizer: 第38回日本糖質学会年会
• [Presentation] 大腸菌の膜タンパク質膜挿入機構解明を目指した蛍光標識MPIase類縁体の合成 (2019)
  • Author(s): 藤川紘樹, 梅川雄一, 鈴木苑実, 西山賢一, 島本啓子
  • Organizer: 日本農芸化学会2019年度大会(東京)
• [Presentation] Syntheses and activities of the functional structures of a glycolipid essential for membrane protein integration (2018)
  • Author(s): Kohki Fujikawa, Sonomi Suzuki, Ryohei Nagase, Shiori Ikeda, Shoko Mori, Kaoru Nomura, Ken-ichi Nishiyama, Keiko Shimamoto
  • Organizer: 29th International Carbohydrate Symposium (ICS)
  • Int'l Joint Research
• [Presentation] 大腸菌膜タンパク質膜挿入に関与する糖脂質MPIase部分構造の合成と活性 (2018)
  • Author(s): 藤川紘樹, 鈴木苑実, 池田汐里, 森祥子, 野村薫, 西山賢一, 島本啓子
  • Organizer: 第37回日本糖質学会年会(仙台)
• [Presentation] 大腸菌の膜タンパク質膜挿入に必須な糖脂質MPIase類縁体の合成と活性 (2018)
  • Author(s): 藤川紘樹, 鈴木苑実, 池田汐里, 森祥子, 野村薫, 西山賢一, 島本啓子
  • Organizer: 第60回天然有機化合物討論会(久留米)
• [Presentation] Syntheses and activities of the functional structures of MPIase, a glycolipid essential for membrane protein integration in E. coli (2018)
  • Author(s): Kohki Fujikawa, Sonomi Suzuki, Ryohei Nagase, Shiori Ikeda, Shoko Mori, Kaoru Nomura, Ken-ichi Nishiyama, Keiko Shimamoto
  • Organizer: The 14th International Kyoto Conference on New Aspects of Organic Chemistry (IKCOC-14)
  • Int'l Joint Research
• [Presentation] 大腸菌膜タンパク質膜挿入に関与する糖脂質MPIase部分構造の合成と活性 (2018)
  • Author(s): 藤川紘樹, 鈴木苑実, 池田汐里, 森祥子, 野村薫, 西山賢一, 島本啓子
  • Organizer: 第5回FCCAシンポジウム・グライコサイエンス若手フォーラム(仙台)
• [Presentation] 大腸菌膜タンパク質の膜挿入機構解明を目指した糖脂質MPIase部分構造の合成 (2017)
  • Author(s): 〇藤川, 西山, 島本
  • Organizer: 日本農芸化学会2017年度大会
• [Presentation] 大腸菌膜タンパク質の膜挿入機構解明を目指した糖脂質MPIase部分構造の合成 (2017)
  • Author(s): 〇藤川, 西山, 島本
  • Organizer: 第16回新規素材探索研究会
• [Presentation] 大腸菌膜タンパク質挿入機構解明を目的とした糖脂質MPIase部分構造の合成 (2017)
  • Author(s): 〇藤川, 池田, 西山, 島本
  • Organizer: 第36回日本糖質学会年会
• [Presentation] 大腸菌膜タンパク質膜挿入機構解明に向けた糖脂質MPIase類縁体の合成と活性 (2017)
  • Author(s): 〇藤川, 池田, 西山, 島本
  • Organizer: 第4回新学術領域研究「反応の集積化が導く中分子戦略」若手シンポジウム
• [Presentation] Syntheses of partial structures of glycolipid MPIase for elucidating the mechanism on membrane protein integration in the inner membrane of E. coli. (2017)
  • Author(s): 〇Fujikawa, Ikeda, Nishiyama, Shimamoto
  • Organizer: International Seminar on Biophysics and Chemical Biology of Biomembrane and Lipid Bilayers
  • Int'l Joint Research
• [Presentation] Syntheses of partial structures of glycolipid MPIase for elucidating the mechanism on membrane protein integration of E. coli. (2017)
  • Author(s): 〇藤川
  • Organizer: 第80回糖鎖科学講演会
  • Invited
• [Book] 化学と生物 (2020)
  • Author(s): 沢里克宏, 藤川紘樹, 島本啓子, 西山賢一
  • Total Pages: 8
  • Publisher: 国際文献社