Molecular dissection of organization and dynamics of membrane lipid domains

• Project/Area Number: 25293015
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: KOBAYASHI Toshihide 国立研究開発法人理化学研究所, 佐甲細胞情報研究室, 客員主管研究員 (60162004)
• Co-Investigator(Kenkyū-buntansha): 村手 源英 国立研究開発法人理化学研究所, その他部局等, 研究員 (30311369) ; 石塚 玲子 国立研究開発法人理化学研究所, その他部局等, 研究員 (60342747) ; 阿部 充宏 国立研究開発法人理化学研究所, その他部局等, 研究員 (90415068) ; 岸本 拓磨 国立研究開発法人理化学研究所, 小林脂質生物学研究室, 基礎科学特別研究員 (70585158)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥19,240,000 (Direct Cost: ¥14,800,000、Indirect Cost: ¥4,440,000); Fiscal Year 2015: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2014: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2013: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000)
• Keywords: 脂質ドメイン / 脂質ラフト / 脂質の可視化 / 脂質結合毒素 / コレステロール / スフィンゴ脂質 / 脂質の非対称性 / スフィンゴミエリン / 脂質プローブ / インフルエンザ / ラマン分光 / カベオレ / ホスホリパーゼC / 脂肪滴 / PLIN2 / Rab18 / フリーズフラクチャー / 脂質結合性毒素 / 上皮細胞 / 細胞分裂 / 形質膜

Outline of Final Research Achievements

In the biomembranes, lipids are not randomly distributed. Rather, lipids are asymmetrically distributed between outer and inner membrane bilayer. Even in one layer, lipids form domains. However, the detailed distribution of lipids in biomembranes are not yet elucidated. We have developed various protein probes that bind specific lipids or lipid domains. Using these probes in combination with state-of-the-art microscopy techniques, we clarified the detailed distribution of lipid domains, including lipid rafts, in biomembranes. Application of our strategy to various pathophysiological events such as cell division, cell adhesion and genetic defects in cholesterol and sphingolipid metabolism provided us new aspects of lipid domains.

Research Products

• [Int'l Joint Research] 国立科学研究センター（CNRS）/国立衛生医学研究所（INSERM）(フランス)
• [Int'l Joint Research] 国立化学研究所(スロベニア)
• [Int'l Joint Research] クイーンズ大学(オーストラリア)
• [Int'l Joint Research] 国立科学研究センター（CNRS）/国立衛生医学研究所（INSERM）(フランス)
• [Int'l Joint Research] クイーンズランド大学(オーストラリア)
• [Int'l Joint Research] 国立化学研究所(スロベニア)
• [Int'l Joint Research] マギル大学(カナダ)
• [Int'l Joint Research] 国立化学研究所/リュブリャナ大学(スロベニア)
• [Int'l Joint Research] クイーンズランド大学(オーストラリア)
• [Int'l Joint Research] ストラスブール大学/国立衛生医学研究所/国立科学センター(フランス)
• [Journal Article] Photoswitchable phospholipid FRET acceptor: Detergent free intermembrane transfer assay of fluorescent lipid analogs. (2017)
  • Author(s): Sumi M, Makino A, Inaba T, Sako Y, Fujimori F, Greimel P, Kobayashi T
  • Journal Title: Sci Rep Volume: in press
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A novel spingomyeling/cholesterol specific probe reveals the dynamics of the membrane domain during virus and in Nieman-Pick type C (2016)
  • Author(s): Makino, A., Sako, Y. et al.
  • Journal Title: FASEB J. Volume: 31 Issue: 4 Pages: 1301-1322
  • DOI: 10.1096/fj.201500075r
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Detection of sphingomyelin clusters by Raman spectroscopy. (2016)
  • Author(s): Shirota K, Yagi K, Inaba T, Li PC, Murata M, Sugita Y, Kobayashi T.
  • Journal Title: Biophys J Volume: 111 Issue: 5 Pages: 999-1007
  • DOI: 10.1016/j.bpj.2016.07.035
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Acute accumulation of free cholesterol induces the degradation of perilipin 2 and Rab18-dependent fusion of ER and lipid droplets in cultured human hepatocytes. (2016)
  • Author(s): Makino A, Hullin-Matsuda F, Murate M, Abe M, Tomishige N, Fukuda M, Yamashita S, Fujimoto T, Vidal H, Lagarde M, Delton I, Kobayashi T.
  • Journal Title: Mol Biol Cell Volume: 27 Issue: 21 Pages: 3293-3304
  • DOI: 10.1091/mbc.e15-10-0730
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Phospholipase Cβ1 induces membrane tubulation and is involved in caveolae formation. (2016)
  • Author(s): Inaba T, Kishimoto T, Murate M, Tajima T, Sakai S, Abe M, Makino A, Tomishige N, Ishitsuka R, Ikeda Y, Takeoka S, Kobayashi T.
  • Journal Title: Proc Natl Acad Sci USA Volume: 113 Issue: 28 Pages: 7834-7839
  • DOI: 10.1073/pnas.1603513113
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Detectors for evaluating the cellular landscape of sphingomyelin- and cholesterol-rich membrane domains (2016)
  • Author(s): Kishimoto T*, Ishitsuka R*, Kobayashia T. * equally contributed
  • Journal Title: Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids Volume: in press Issue: 8 Pages: 812-829
  • DOI: 10.1016/j.bbalip.2016.03.013
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Pore-forming toxins: Properties, diversity, and uses as tools to image sphingomyelin and ceramide phosphoethanolamine. (2016)
  • Author(s): Yamaji-Hasegawa A, Hullin-Matsuda F, Greimel P, Kobayashi T.
  • Journal Title: Biochim. Biophys Acta Volume: 1858 Issue: 3 Pages: 576-592
  • DOI: 10.1016/j.bbamem.2015.10.012
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Visualization of lipid membrane reorganization induced by a pore-forming toxin using high-speed atomic force microscopy (2015)
  • Author(s): Yilmaz N, Kobayashi T
  • Journal Title: ＡＣＳ Ｎａｎｏ Volume: 9 Issue: 8 Pages: 7960-7967
  • DOI: 10.1021/acsnano.5b01041
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Asymmetric diacylglycerol dynamics on the cytoplasmic and luminal sides of a single endomembrane in living cells (2015)
  • Author(s): Ueda Y, Ogiso H, Sato M, Umezawa Y, Okazaki T, Kobayashi T
  • Journal Title: Ｓｃｉ Ｒｅｐ Volume: 5 Issue: 1 Pages: 12960-12960
  • DOI: 10.1038/srep12960
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Evaluation of aegerolysins as novel tools to detect and visualize ceramide phosphoethanolamine, a major sphingolipid in invertebrates (2015)
  • Author(s): Bhat HB, Ishitsuka R, Inaba T, Murate M, Abe M, Makino A, Kohyama-Koganeya A, Nagao K, Kurahashi A, Kishimoto T, Tahara M, Yamano A, Nagamune K, Hirabayashi Y, Juni N, Umeda M, Fujimori F, Nishibori K, Yamaji-Hasegawa A, Greimel P, Kobayashi T.
  • Journal Title: FASEB J Volume: 29(9) Issue: 9 Pages: 3920-34
  • DOI: 10.1096/fj.15-272112
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Transbilayer distribution of lipids at nano scale. (2015)
  • Author(s): Murate M, Abe M, Kasahara K, Iwabuchi K, Umeda M, Kobayashi T
  • Journal Title: J Cell Sci Volume: 128 Pages: 1627-1638
  • DOI: 10.1242/jcs.163105
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Visualization of the heterogeneous distribution of sphingomyelin associated with cytokinesis, cell polarity and sphingolipidosis (2015)
  • Author(s): Makino A, Abe M, Murate M, Inaba T, Yilmaz N, Hullin-Matsuda F, Kishimoto T, Schieber NL, Taguchi T, Arai H, Anderluh G, Parton RG, Kobayashi T.
  • Journal Title: FASEB J Volume: 29 Issue: 2 Pages: 477-493
  • DOI: 10.1096/fj.13-247585
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Lipid compartmentalization in the endosome system (2014)
  • Author(s): Hullin-Matsuda F, Taguchi T, Greimel P, Kobayashi T.
  • Journal Title: Sem Cell Dev Biol Volume: (in press) Pages: 48-56
  • DOI: 10.1016/j.semcdb.2014.04.010
  • Peer Reviewed
• [Journal Article] Real-Time Visualization of Assembling of a Sphingomyelin-Specific Toxin. (2013)
  • Author(s): N. Yilmaz, T. Yamada, P. Greimel, T. Uchihashi, T. Ando, T. Kobayashi
  • Journal Title: Biophys. J. Volume: 105 Issue: 6 Pages: 1397-1405
  • DOI: 10.1016/j.bpj.2013.07.052
  • Peer Reviewed
• [Presentation] A novel sphingomyelin/cholesterol domain-specific probe reveals the dynamics of the membrane domains during virus release (2016)
  • Author(s): Makino A, Abe M, Ishitsuka R, Murate M, Kishimoto T, Sakai S, Hullin-Matsuda F, Inaba T, Greimel P, Kobayashi T
  • Organizer: 12th Gerli international lipidomics meeting
  • Place of Presentation: Toulouse, France
  • Year and Date: 2016-10-23
  • Int'l Joint Research
• [Presentation] Phosphatidylglycerol analog regulates the synthesis of the endosome-specific and fusion enhancing lipid bis(monoacylglycero)phosphate (2016)
  • Author(s): Tan HH, Hulin-Matsuda F, Lim SR, Sudesh K, Gam LH, Greimel P, Kobayashi T
  • Organizer: 12th Gerli international lipidomics meeting
  • Place of Presentation: Toulouse, France
  • Year and Date: 2016-10-23
  • Int'l Joint Research
• [Presentation] Transbilayer distribution of lipids in the plasma membrane revealed by immuno-electron microscopy (2016)
  • Author(s): Murate M
  • Organizer: 57th International Conference on the Bioscience of Lipids
  • Place of Presentation: Chamonix Mont-Blanc, France
  • Year and Date: 2016-09-04
  • Int'l Joint Research
• [Presentation] Elucidation of the natural regioisomer of the endosome-specific lipid bis(monoacylglycero)phosphate (2016)
  • Author(s): Lim SR, Tan HH, Hullin-Matsuda F, Sudesh K, Gram LH, Greimel P, Kobayashi T
  • Organizer: 57th International Conference on the Bioscience of Lipids
  • Place of Presentation: Chamonix Mont-Blanc, France
  • Year and Date: 2016-09-04
  • Int'l Joint Research
• [Presentation] Phosphatidylglycerol analog regulates the synthesis of the endosome-specific lipid bis(monoacylglycero)phosphate (2016)
  • Author(s): Tan HH, Hullin-Matsuda F, Lim SR, Sudesh K, Gram LH, Greimel P, Kobayashi T
  • Organizer: 57th International Conference on the Bioscience of Lipids
  • Place of Presentation: Chamonix Mont-Blanc, France
  • Year and Date: 2016-09-04
  • Int'l Joint Research
• [Presentation] Phospholipase C beta 1 induces membrane tubulation and is involved in caveolae formation (2016)
  • Author(s): Inaba T, Kishimoto T, Murate M, Tajima T, Sakai S, Abe M, Makino A, Tomishige N, Ishitsuka R, Ikeda Y, Takeoka S, Kobayashi T
  • Organizer: 57th International Conference on the Bioscience of Lipids
  • Place of Presentation: Chamonix Mont-Blanc, France
  • Year and Date: 2016-09-04
  • Int'l Joint Research
• [Presentation] Imaging lipids and lipid domains (2016)
  • Author(s): Kobayashi T
  • Organizer: Joint meeting of the membrane sections of the French and German Biophysical Societies
  • Place of Presentation: Bad Herrenalb, Germany
  • Year and Date: 2016-04-11
  • Int'l Joint Research
• [Presentation] Cellular localization of sphingomyelin clusters (2016)
  • Author(s): Kobayashi T
  • Organizer: Gordon Research Conference on Glycolipid and Sphingolipid Biology
  • Place of Presentation: Lucca, Italy
  • Year and Date: 2016-03-06
  • Int'l Joint Research / Invited
• [Presentation] Imaging lipids and lipid domains (2016)
  • Author(s): Kobayashi T
  • Organizer: Sphingonet’s International Colloquium
  • Place of Presentation: Amsterdam, Netherland
  • Year and Date: 2016-01-27
  • Int'l Joint Research / Invited
• [Presentation] Imaging lipids and lipid domains (2015)
  • Author(s): Kobayashi T
  • Organizer: 11th GERLI Lipidomics Meeting
  • Place of Presentation: Strasbourg France
  • Year and Date: 2015-10-25
  • Int'l Joint Research / Invited
• [Presentation] 膜を横切る脂質の運動、フリップフロップについて (2013)
  • Author(s): 小林俊秀
  • Organizer: 第10回「生体膜・コロイド研究の最前線」
  • Place of Presentation: 東海村
  • Invited
• [Presentation] Free cholesterol loading induces the degradation of perilipin 2 and rab18-de pendent abnormal lipidation of apolipoprotein B in cultured hepatocytes (2013)
  • Author(s): Makino A, Hullin-Matsuda F, Murate M, Abe M, Fukuda M, Yamashita S, Fujimoto T, Vidal H, Lagarde M, Delton-Vandenbroucke I, Kobayashi T
  • Organizer: 第８６回日本生化学会大会
  • Place of Presentation: 横浜
  • Invited
• [Presentation] Imaging lipids and lipid domains
  • Author(s): Kobayashi T
  • Organizer: 2nd Mini Symposium “Protein-lipid interactions”
  • Place of Presentation: Piran, Slovenia
  • Invited
• [Presentation] 脂質のイメージング
  • Author(s): 小林俊秀
  • Organizer: 第1回JFAS講演会
  • Place of Presentation: 秋葉原
  • Invited
• [Remarks]
  • URL: http://lipidology.riken.jp/
• [Remarks]
  • URL: http://lipidology.riken.jp/