蛍光標識脂質を用いた脂質膜の動的秩序解析

• Project Area: Dynamical ordering of biomolecular systems for creation of integrated functions
• Project/Area Number: 16H00773
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: Kyushu University
• Principal Investigator: 松森 信明 九州大学, 理学研究院, 教授 (50314357)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2017: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000); Fiscal Year 2016: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
• Keywords: 脂質ラフト / スフィンゴミエリン / セラミド / 蛍光

Outline of Annual Research Achievements

脂質ラフトはコレステロールやスフィンゴ脂質に富んだ秩序性の高い領域で、近年その生理的重要性が認識されてきた。一方で、生成と崩壊を繰り返しつつ自己組織化しているため、特に生細胞膜におけるラフト形成の分子基盤は未解明のままである。我々は本新学術の第一期において時間分解能の高い蛍光寿命測定や共焦点蛍光顕微鏡を導入し、ラフト観察に成果を挙げてきた。本研究では、第一期において有用性が示された蛍光スフィンゴミエリン(SM)の設計脂質に基づいて、新たに各種脂質の蛍光標識体を開発し、蛍光観察による脂質膜動的秩序改正の高度化を図る。さらに、単純な人工膜ばかりでなく、脂質の外葉と内葉の組成の異なる非対称膜、さらには生細胞膜へと研究対象を拡大していくことを目指した。
本研究では、すでにSM、およびSMのトランス二重結合を還元したジヒドロスフィンゴミエリンの蛍光標識体の調製に成功した。またアシル鎖長の異なるSM蛍光標識体も合成し、ラフトにおける挙動の解析を行ったところ、予想に反して長鎖のSMのラフト集積性が低くなる結果が得られた（論文準備中）。さらにセラミドに蛍光基を導入した蛍光セラミドの合成にも成功し、論文発表およびプレスリリースを行った（Langmuir 2019）。これらの標識体はいずれももとの脂質の性質をよく再現していた。さらにラフトの可視化を目指し、蛍光SM間のFRETによりラフトをより鮮明に可視化する技術の開発にも成功した（Chem Phys Lipids 2018）。これらの蛍光脂質はすでに生体膜にも導入し、ラフトの可視化および脂質の動的秩序の解明に成功した（Sci Rep.2017)。さらに非対称膜の作成にも成功し、外葉に形成された脂質ラフトが内葉に与える影響について蛍光脂質を用いて解析した。このように当初予定していた以上の研究成果を達成することができた。

Research Progress Status

平成29年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

平成29年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] Abo Akademi University(フィンランド)
• [Journal Article] A concise method for quantitative analysis of interactions between lipids and membrane proteins (2019)
  • Author(s): Inada Masataka、Kinoshita Masanao、Sumino Ayumi、Oiki Shigetoshi、Matsumori Nobuaki
  • Journal Title: Analytica Chimica Acta Volume: 1059 Pages: 103-112
  • DOI: 10.1016/j.aca.2019.01.042
  • Peer Reviewed / Open Access
• [Journal Article] Preparation and Membrane Distribution of Fluorescent Derivatives of Ceramide (2019)
  • Author(s): Matsufuji Takaaki、Kinoshita Masanao、Matsumori Nobuaki
  • Journal Title: Langmuir Volume: 35 Issue: 6 Pages: 2392-2398
  • DOI: 10.1021/acs.langmuir.8b03176
  • Peer Reviewed
• [Journal Article] Theonellamide A, a marine-sponge-derived bicyclic peptide, binds to cholesterol in aqueous DMSO: Solution NMR-based analysis of peptide-sterol interactions using hydroxylated sterol (2019)
  • Author(s): Cornelio Kimberly、Espiritu Rafael Atillo、Hanashima Shinya、Todokoro Yasuto、Malabed Raymond、Kinoshita Masanao、Matsumori Nobuaki、Murata Michio、Nishimura Shinichi、Kakeya Hideaki、Yoshida Minoru、Matsunaga Shigeki
  • Journal Title: Biochimica et Biophysica Acta (BBA) - Biomembranes Volume: 1861 Issue: 1 Pages: 228-235
  • DOI: 10.1016/j.bbamem.2018.07.010
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] On the Importance of the C(1)-OH and C(3)-OH Functional Groups of the Long-Chain Base of Ceramide for Interlipid Interaction and Lateral Segregation into Ceramide-Rich Domains (2018)
  • Author(s): Mouts Anna、Vattulainen Elina、Matsufuji Takaaki、Kinoshita Masanao、Matsumori Nobuaki、Slotte J. Peter
  • Journal Title: Langmuir Volume: 34 Issue: 51 Pages: 15864-15870
  • DOI: 10.1021/acs.langmuir.8b03237
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Sphingomyelin Stereoisomers Reveal That Homophilic Interactions Cause Nanodomain Formation (2018)
  • Author(s): Yano Yo、Hanashima Shinya、Yasuda Tomokazu、Tsuchikawa Hiroshi、Matsumori Nobuaki、Kinoshita Masanao、Al Sazzad Md. Abdullah、Slotte J. Peter、Murata Michio
  • Journal Title: Biophysical Journal Volume: 115 Issue: 8 Pages: 1530-1540
  • DOI: 10.1016/j.bpj.2018.08.042
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Evidence of lipid rafts based on the partition and dynamic behavior of sphingomyelins (2018)
  • Author(s): Kinoshita Masanao、Suzuki Kenichi G.N.、Murata Michio、Matsumori Nobuaki
  • Journal Title: Chemistry and Physics of Lipids Volume: 215 Pages: 84-95
  • DOI: 10.1016/j.chemphyslip.2018.07.002
  • Peer Reviewed
• [Journal Article] Synthesis and Stereochemical Revision of the C31-C67 Fragment of Amphidinol 3 (2018)
  • Author(s): Wakamiya Yuma、Ebine Makoto、Murayama Mariko、Omizu Hiroyuki、Matsumori Nobuaki、Murata Michio、Oishi Tohru
  • Journal Title: Angewandte Chemie Volume: 130 Issue: 21 Pages: 6168-6172
  • DOI: 10.1002/ange.201712167
  • Peer Reviewed
• [Journal Article] Synthesis and Complete Structure Determination of a Sperm-Activating and -Attracting Factor Isolated from the Ascidian Ascidia sydneiensis (2018)
  • Author(s): Watanabe Tomohiro、Shibata Hajime、Ebine Makoto、Tsuchikawa Hiroshi、Matsumori Nobuaki、Murata Michio、Yoshida Manabu、Morisawa Masaaki、Lin Shu、Yamauchi Kosei、Sakai Ken、Oishi Tohru
  • Journal Title: Journal of Natural Products Volume: 81 Issue: 4 Pages: 985-997
  • DOI: 10.1021/acs.jnatprod.7b01052
  • Peer Reviewed
• [Journal Article] Dynamic membrane interactions of antibacterial and antifungal biomolecules, and amyloid peptides, revealed by solid-state NMR spectroscopy (2018)
  • Author(s): Naito Akira、Matsumori Nobuaki、Ramamoorthy Ayyalusamy
  • Journal Title: Biochimica et Biophysica Acta (BBA) - General Subjects Volume: 1862 Issue: 2 Pages: 307-323
  • DOI: 10.1016/j.bbagen.2017.06.004
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Recent Solid-State NMR Studies of Hydrated Lipid Membranes (2018)
  • Author(s): Umegawa Yuichi、Matsumori Nobuaki、Murata Michio
  • Journal Title: Annu. Rep. NMR Spectrosc. Volume: 94 Pages: 41-72
  • DOI: 10.1016/bs.arnmr.2017.12.003
  • Peer Reviewed
• [Journal Article] 蛍光修飾したスフィンゴミエリン誘導体の脂質ラフト観察への応用 (2018)
  • Author(s): 木下 祥尚, 松森信明
  • Journal Title: 生物物理 Volume: 58 Pages: 321-323
  • NAID: 130007522421
• [Journal Article] 生体膜を標的にする天然有機化合物 (2018)
  • Author(s): 西村 慎一, 掛谷 秀昭, 松森信明
  • Journal Title: 化学と生物 Volume: 56 Pages: 678-685
• [Journal Article] Preparation and Membrane Properties of Oxidized Ceramide Derivatives (2017)
  • Author(s): Matsufuji Takaaki、Kinoshita Masanao、Mouts Anna、Slotte J. Peter、Matsumori Nobuaki
  • Journal Title: Langmuir Volume: 34 Issue: 1 Pages: 465-471
  • DOI: 10.1021/acs.langmuir.7b02654
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] NMR Studies on Natural Product-Stereochemical Determination and Conformational Analysis in Solution and in Membrane (2017)
  • Author(s): Matsumori Nobuaki、Murata Michio
  • Journal Title: Experimental Approaches of NMR Spectroscopy Volume: - Pages: 383-414
  • DOI: 10.1007/978-981-10-5966-7_14
  • ISBN: 9789811059650, 9789811059667
  • Peer Reviewed
• [Journal Article] The impact of metal complex lipids on viscosity and curvature of hybrid liposomes (2017)
  • Author(s): R. Ohtani, T. Tokita, T. Takaya, K. Iwata, M. Kinoshita, N. Matsumori, M. Nakamura, L. F. Lindoy, S. Hayami
  • Journal Title: Chem. Commun. Volume: 53 Issue: 99 Pages: 13249-13252
  • DOI: 10.1039/c7cc07944c
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Structures of the Largest Amphidinol Homologues from the Dinoflagellate Amphidinium carterae and Structure-Activity Relationships (2017)
  • Author(s): M. Satake, K. Cornelio, S. Hanashima, R. Malabed, M. Murata, N. Matsumori, H. Zhang, F. Hayashi, S. Mori, J. S. Kim, C.-H. Kim, J.-S. Lee
  • Journal Title: Journal of Natural Products Volume: 80 Issue: 11 Pages: 2883-2888
  • DOI: 10.1021/acs.jnatprod.7b00345
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Channel Formation and Membrane Deformation via Sterol-Aided Polymorphism of Amphidinol 3 (2017)
  • Author(s): Iwamoto M, Sumino A, Shimada E, Kinoshita M, Matsumori N, Oiki S
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 10782-10782
  • DOI: 10.1038/s41598-017-11135-x
  • Peer Reviewed / Open Access
• [Journal Article] Emphatic visualization of sphingomyelin-rich domains by inter-lipid FRET imaging using fluorescent sphingomyelins (2017)
  • Author(s): 1.Kinoshita M, Ano H, Murata M, Shigetomi K, Ikenouchi J, Matsumori N
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 16801-16801
  • DOI: 10.1038/s41598-017-16361-x
  • Peer Reviewed / Open Access
• [Journal Article] Raft-based sphingomyelin interactions revealed by new fluorescent sphingomyelin analogs (2017)
  • Author(s): Masanao Kinoshita, Kenichi GN Suzuki, Nobuaki Matsumori, Misa Takada, Hikaru Ano, Kenichi Morigaki, Mitsuhiro Abe, Asami Makino, Toshihide Kobayashi, Koichiro M Hirosawa, Takahiro K Fujiwara, Akihiro Kusumi, Michio Murata
  • Journal Title: J Cell Biol Volume: 未定 Issue: 4 Pages: 1183-1204
  • DOI: 10.1083/jcb.201607086
  • NAID: 120005998045
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Membrane protein structure determination by SAD, SIR, or SIRAS phasing in serial femtosecond crystallography using an iododetergent (2016)
  • Author(s): Takanori Nakane, Shinya Hanashima, Mamoru Suzuki, Haruka Saiki, Taichi Hayashi, Keisuke Kakinouchi, Shigeru Sugiyama, Satoshi Kawatake, Shigeru Matsuoka, Nobuaki Matsumori, et al
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 113 Issue: 46 Pages: 13039-13044
  • DOI: 10.1073/pnas.1602531113
  • Peer Reviewed / Open Access
• [Journal Article] Sterol-dependent membrane association of the marine sponge-derived bicyclic peptide Theonellamide A as examined by 1H NMR (2016)
  • Author(s): Kimberly Cornelio, Rafael Atillo Espiritu, Yasuto Todokoro, Shinya Hanashima, Masanao Kinoshita, Nobuaki Matsumori, Michio Murata, Shinichi Nishimura, Hideaki Kakeya, Minoru Yoshida, Shigeki Matsunaga
  • Journal Title: Bioorganic & Medicinal Chemistry Volume: 24 Issue: 21 Pages: 5235-5242
  • DOI: 10.1016/j.bmc.2016.08.043
  • Peer Reviewed
• [Journal Article] Marine sponge cyclic peptide theonellamide A disrupts lipid bilayer integrity without forming distinct membrane pores. (2016)
  • Author(s): 1.Espiritu, R. A., Cornelio, K., Kinoshita, M., Matsumori, N., Murata, M., Nishimura, S., Kakeya, H., Yoshida, M. & Matsunaga, S.
  • Journal Title: Biochim. Biophys. Acta Volume: 1858 Issue: 6 Pages: 1373-1379
  • DOI: 10.1016/j.bbamem.2016.03.019
  • Peer Reviewed
• [Journal Article] The Structure of the Bimolecular Complex between Amphotericin B and Ergosterol in Membranes Is Stabilized by Face-to-Face van der Waals Interaction with Their Rigid Cyclic Cores (2016)
  • Author(s): Yasuo Nakagawa, Yuichi Umegawa, Naohiro Matsushita, Tomoya Yamamoto, Hiroshi Tsuchikawa, Shinya Hanashima, Tohru Oishi, Nobuaki Matsumori, Michio Murata
  • Journal Title: Biochemistry Volume: 55 Issue: 24 Pages: 3392-3402
  • DOI: 10.1021/acs.biochem.6b00193
  • Peer Reviewed
• [Journal Article] Lipid Interactions and Organization in Complex Bilayer Membranes (2016)
  • Author(s): Engberg, O; Yasuda, T; Hautala, V; Matsumori, N; Nyholm, TKM; Murata, M; Slotte, JP
  • Journal Title: BIOPHYSICAL JOURNAL Volume: 110 Issue: 7 Pages: 1563-1573
  • DOI: 10.1016/j.bpj.2015.12.043
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 脂質間FRETを用いたコントラストの強調とスフィンゴミエリンリッチ領域の可視化 (2019)
  • Author(s): 木下祥尚，阿野光，重富健太，村田道雄，池内順一，松森信明
  • Organizer: 日本化学会第99回春季年会2019
• [Presentation] 低流量電子線散乱法を用いた局所的な脂質充填構造の解析 (2019)
  • Author(s): 山口晋平，木下祥尚， 松森信明
  • Organizer: 日本化学会第99回春季年会2019
• [Presentation] アシル鎖長の異なるスフィンゴミエリンがドメイン形成に与える影響ー蛍光スフィンゴミエリンを用いた検討 (2019)
  • Author(s): 平野佳奈，木下祥尚， 松森信明
  • Organizer: 日本化学会第99回春季年会2019
• [Presentation] 脂質膜内葉と外葉の非対称性が人工ラフト膜の構造や物性に与える影響 (2019)
  • Author(s): 古閑卓也，木下祥尚，松森信明
  • Organizer: 日本化学会第99回春季年会2019
• [Presentation] 脂質固定ビーズによる脂質特異的タンパク質の同定 (2019)
  • Author(s): 安田裕貴， 松藤嵩明，木下祥尚，松森信明
  • Organizer: 日本化学会第99回春季年会2019
• [Presentation] 人工膜脂質に対する吸入麻酔剤の作用 (2019)
  • Author(s): 川口翼，木下祥尚，松森信明
  • Organizer: 日本化学会第99回春季年会2019
• [Presentation] Detailed insight for the effect of specific interaction with lipids on membrane protein activation (2019)
  • Author(s): Masataka Inada，Masanao Kinoshita，Masayuki Iwamoto，Shigetoshi Oiki，Nobuaki Matsumori
  • Organizer: 日本化学会第99春季年会
• [Presentation] Crucial Role of Specific Interactions with Archeal Glycolipid S-TGA-1 in Structuralization and Functionalization of Bacteriorhodopsin (2018)
  • Author(s): Masataka Inada，Masanao Kinoshita，Nobuaki Matsumori
  • Organizer: 第56回日本生物物理学会年会
• [Presentation] 膜タンパク質-脂質相互作用に対する定量評価法の開発および特異的脂質の同定 (2018)
  • Author(s): 稲田壮峰，木下祥尚，松森信明
  • Organizer: 日本分析化学会第67年会
• [Presentation] 異なるアシル鎖をもつ蛍光スフィンゴミエリの調製とドメイン形成の解析 (2018)
  • Author(s): 平野佳奈，木下祥尚，松森信明
  • Organizer: 第12回バイオ関連化学シンポジウム
• [Presentation] 脂質膜中における膜作用性海洋天然物アンフィジノール3とステロールの相互作用 (2018)
  • Author(s): 檜枝愛美， 木下祥尚，松森信明
  • Organizer: 第12回バイオ関連化学シンポジウム
• [Presentation] Development of a concise method for quantitative analysis of interactions between membrane proteins (2018)
  • Author(s): Masataka Inada，Masanao Kinoshita，Nobuaki Matsumori
  • Organizer: International Workshop on Biological Membranes : Tiny Lipids With Grand Function
  • Int'l Joint Research
• [Presentation] NMR and Fluorescent Studies of Lipid Rafts (2016)
  • Author(s): Nobuaki Matsumori and Masanao Kinoshita
  • Organizer: 9th Korea-Japan Seminars on Biomolecular Sciences: Experimental and Simulations
  • Place of Presentation: Gyeongju, Korea
  • Int'l Joint Research / Invited
• [Book] 天然物の化学Ⅱ（科学のとびら64) (2018)
  • Author(s): 上村 大輔 編
  • Total Pages: 192
  • Publisher: 東京化学同人
  • ISBN: 9784807915057
• [Book] Experimental Approaches of NMR Spectroscopy. Methodology and Application to Life Science and Materials Science (2017)
  • Author(s): Naito, Akira 編
  • Total Pages: 636
  • Publisher: Springer
  • ISBN: 9789811059650
• [Remarks] 九州大学大学院理学研究院化学
  • URL: http://www.scc.kyushu-u.ac.jp/BioanalChem/index.html
• [Remarks] 九州大学大学院理学研究院化学部門生体分析化学研究室
  • URL: http://www.scc.kyushu-u.ac.jp/BioanalChem/