Molecular dissection of the organization and dynamics of lipid domains in biomembranes

• Project/Area Number: 22390018
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: KOBAYASHI Toshihide 独立行政法人理化学研究所, 小林脂質生物学研究室, 主任研究員 (60162004)
• Co-Investigator(Kenkyū-buntansha): ISHITSUKA Reiko 独立行政法人理化学研究所, 小林脂質生物学研究室, 専任研究員 (60342747) ; ABE Mitsuhiro 独立行政法人理化学研究所, 小林脂質生物学研究室, 研究員 (90415068) ; MURATE Motohide 独立行政法人理化学研究所, 小林脂質生物学研究室, 研究員 (30311369) ; MAKINO Asami 独立行政法人理化学研究所, 小林脂質生物学研究室, 特別研究員 (20373368)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000); Fiscal Year 2012: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2011: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2010: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000)
• Keywords: 脂質ドメイン / 脂質プローブ / 脂質ラフト / 脂質の非対称性 / スフィンゴミエリン / コレステロール / 脂質の可視化 / ホスファチジルイノシトール4,5-ビスリン酸 / 細胞分裂 / 超高解像顕微鏡 / スフィンゴ脂質 / 免疫電子顕微鏡 / X線散乱

Research Abstract

We established a method to visualize major components of lipid rafts, sphingomyelin and cholesterol, in sub-micron scale.
New screening protocol using lipid imaging was employed to isolate a new inhibitor of sphingomyelin biosynthesis.
The role of sphingomyelin to regulate the transbilayer movement of lipid second messengers such as ceramide and diacylglycerol was elucidated.
The unusual stereoconfiguration of endosome-specific lipid was determined.

Research Products

• [Journal Article] Sphingomyelin regulates the transbilayer movement of diacylglycerol in the plasma membrane of Madin-Darby canine kidney cells (2013)
  • Author(s): Ueda Y, Makino A, Murase-Tamada K, Sakai S, Inaba T, Hullin-Matsuda F, and T. Kobayashi.
  • Journal Title: FASEB J Volume: (in press)
  • Peer Reviewed
• [Journal Article] Role for phospholipid flippase complex of ATP8A1 and CDC50A proteins in cell migration (2013)
  • Author(s): Kato U, Inadome H, Yamamoto M, Emoto K, Kobayashi T, and M. Umeda
  • Journal Title: J Biol Chem Volume: 288 Pages: 4922-4934
  • Peer Reviewed
• [Journal Article] Role of phospholipid flippase complex ATP8A1 and CDC50A proteins in cell migration (2013)
  • Author(s): Utako Kato
  • Journal Title: J Biol Chem Volume: 288 Pages: 4922-4934
  • Peer Reviewed
• [Journal Article] Limonoid compounds inhibit sphingomyelin biosynthesis by preventing CERT protein-dependent extraction of ceramides from the endoplasmic reticulum (2012)
  • Author(s): Hullin-Matsuda F, Tomishige N, Sakai S, Ishitsuka R, Ishii K, Makino A, Greimel P, Abe M, Laviad EL, Lagarde M, Vidal H, Saito T, Osada H, Hanada K, Futerman AH, and T. Kobayashi
  • Journal Title: J Biol Chem Volume: 287 Pages: 24397-24411
  • Peer Reviewed
• [Journal Article] A role for sphingomyelin-rich lipid domains in the accumulation of PIP2 to the cleavage furrow during cytokinesis (2012)
  • Author(s): Abe M, Makino A, Hullin-Matsuda F, Kamijo K, Ohno-Iwashita Y, Hanada K, Mizuno H, Miyawaki A, and T. Kobayashi
  • Journal Title: Mol Cell Biol Volume: 32 Pages: 1396-1407
  • Peer Reviewed
• [Journal Article] Nano-mechanical recognition of sphingomyelin-rich membrane domains by atomic force microscopy (2012)
  • Author(s): Wang T, Shogomori H, Hara M, Yamada T, and T. Kobayashi
  • Journal Title: Biochemistry Volume: 51 Pages: 74-82
  • Peer Reviewed
• [Journal Article] Rapid flip-flop motions of diacylglycerol and ceramide in phospholipid bilayers (2012)
  • Author(s): Ogushi F, Ishitsuka R, Kobayashi T, and Y. Sugita
  • Journal Title: Chem Phys Lett Volume: 522 Pages: 96-102
  • Peer Reviewed
• [Journal Article] Spectroscopic evidence for the unusual stereochemical configuration of an endosome-specific lipid (2012)
  • Author(s): Tan H-H, Makino A, Sudesh K, Greimel P, and T. Kobayashi
  • Journal Title: Angew Chem Int Ed Volume: 51 Pages: 533-535
  • Peer Reviewed
• [Journal Article] Subcellular localization of sphingomyelin revealed by two toxin-based probes in mammalian cells (2012)
  • Author(s): Rieko Yachi
  • Journal Title: Genes Cells Volume: 17 Pages: 720-727
  • Peer Reviewed
• [Journal Article] The single-giant unilamellar vesicle method reveals lysenin-induced pore formation in lipid membranes containing sphingomyelin (2012)
  • Author(s): Jahangir Md. Alam
  • Journal Title: Biiochemistry Volume: 51 Pages: 5160-5172
  • Peer Reviewed
• [Journal Article] A role for sphingomyelin-rich lipid domains in the accumulation of phosphatidylinositol-4,5-bisphosphate to the cleavage furrow during cytokinesis (2012)
  • Author(s): Mitsuhiro Abe
  • Journal Title: Mol Cell Biol Volume: 32 Pages: 1396-1407
  • Peer Reviewed
• [Journal Article] Lipid polarity is maintained in the absence of tight junctions (2012)
  • Author(s): Ikenouchi J, et al
  • Journal Title: J Biol Chem Volume: (In press)
  • Peer Reviewed
• [Journal Article] A role for sphingomyelin-rich lipid domains in the accumulation of PIP2 to the cleavage furrow during cytokinesis (2012)
  • Author(s): Abe M, et al
  • Journal Title: Mol Cell Biol Volume: (In press)
  • Peer Reviewed
• [Journal Article] Rapid flip-flop motions of diacylglycerol and ceramide in phospholipid bilayers (2012)
  • Author(s): Ogushi, et al
  • Journal Title: Chem Phys Lett Volume: (In press)
  • Peer Reviewed
• [Journal Article] Spectroscopic evidence for the unusual stereochemical configuration of an endosome-specific lipid (2012)
  • Author(s): Tan, et al
  • Journal Title: Angew Chem.Int.Ed.Engl. Volume: 51 Pages: 533-535
  • Peer Reviewed
• [Journal Article] Fluorescence image screening for chemical compounds modifying cholesterol metabolism and distribution (2011)
  • Author(s): Ishitsuka R, Saito T, Osada H, Ohno-Iwashita Y, and T. Kobayashi
  • Journal Title: J Lipid Res. Volume: 52 Pages: 2084-2094
  • Peer Reviewed
• [Journal Article] Fluorescent probes for superresolution imaging of lipid domains on the plasma membrane (2011)
  • Author(s): Mizuno H, Abe M, Dedecker P, Makino A, Rocha S, Ohno-Iwashita Y, Hofkens J, Kobayashi T, and A. Miyawaki
  • Journal Title: Chem Sci Volume: 2 Pages: 1548-1553
  • Peer Reviewed
• [Journal Article] Multiplex analysis of sphingolipids using amine-reactive tags (iTRAQ) (2011)
  • Author(s): Nabetani T, Makino A, Hullin-Matsuda F, Hirakawa T, Takeoka S, Okino N, Ito M, Kobayashi T, and Y. Hirabayashi
  • Journal Title: J Lipid Res Volume: 52 Pages: 1294-1302
  • Peer Reviewed
• [Journal Article] Fluorescence image screening for chemical compounds modifying cholesterol metabolism and distribution (2011)
  • Author(s): Ishitsuka, et al
  • Journal Title: J Lipid Res Volume: 52 Pages: 2084-2094
  • Peer Reviewed
• [Journal Article] SMS1-generated sphingomyelin plays an important role in transferrin trafficking and cell proliferation (2011)
  • Author(s): Shakor, et al
  • Journal Title: J Biol Chem Volume: 286 Pages: 36053-36062
  • Peer Reviewed
• [Journal Article] Phosphatidylglucoside forms specific lipid domains on the outer leaflet of the plasma membrane (2010)
  • Author(s): Murate M, Hayakawa T, Ishii K, Inadome H, Greimel P, Watanabe Y, Nagatsuka Y, Ito K, Ito Y, Takahashi H, Hirabayashi Y, and T. Kobayashi
  • Journal Title: Biochemistry Volume: 49 Pages: 4732-4739
  • Peer Reviewed
• [Journal Article] Marine antifungal theonellamides target 3β-hydroxysterol to activate Rho1 signaling (2010)
  • Author(s): Nishimura S, Arita Y, Honda M, Iwamoto K, Matsuyama A, Shirai A, Kawasaki H, Kakeya H,Kobayashi T, Matsunaga S, and M. Yoshida
  • Journal Title: Nat Chem Biol Volume: 6 Pages: 519-526
  • Peer Reviewed
• [Journal Article] Phosphatidylglucoside forms specific lipid domains on the outer leaflet of the plasma membrane (2010)
  • Author(s): Murate, M.
  • Journal Title: Biochemistry Volume: 49 Pages: 4732-9
  • Peer Reviewed
• [Journal Article] Marine antifungal theonellamides target 3β-hydroxysterol to activate Rho1 signaling (2010)
  • Author(s): Nishimura, S.
  • Journal Title: Nat Chem Biol Volume: 6 Pages: 519-26
  • Peer Reviewed
• [Journal Article] Duramycin-induced destabilization of a phosphatidylethanolamine monolayer at the air-water interface observed by vibrational sum-frequency generation spectroscopy (2010)
  • Author(s): Rzeznicka, I.I.
  • Journal Title: Langmuir Volume: 26 Pages: 16055-62
  • Peer Reviewed
• [Presentation] Protein probes for studying lipid domains (2013)
  • Author(s): Toshihide Kobayashi
  • Organizer: The fourteenth international membrane research forum
  • Place of Presentation: 京都
  • Invited
• [Presentation] 細胞分裂時におけるホスファチジルイノシトール4,5-二リン酸の分裂溝への局在に対するスフィンゴミエリンドメインの役割 (2013)
  • Author(s): 小林俊秀
  • Organizer: 第９０回日本生理学会
  • Place of Presentation: 東京
  • Invited
• [Presentation] 細胞質分裂におけるスフィンゴミエリンドメインの役割 (2012)
  • Author(s): 阿部充宏、牧野麻美、ウラン松田フランソワーズ、小林俊秀
  • Organizer: 第85回生化学会大会
  • Place of Presentation: 福岡
• [Presentation] A role for sphingomyelin-rich lipid domains in the accumulation of phosphatidylinositol-4,5-bisphosphate to the cleavage furrow during cytokinesis (2012)
  • Author(s): Abe, M., A. Makino, F. Hullin-Matsuda, and T. Kobayashi
  • Organizer: Lipid-Protein Interactions in Membranes: Implications for Health and Disease
  • Place of Presentation: Hyderabad, India
• [Presentation] エンドソーム特異的脂質の構造、代謝、機能 (2012)
  • Author(s): 小林俊秀
  • Organizer: 第54回日本脂質生化学会
  • Place of Presentation: 福岡
• [Presentation] Session chair's overview (2012)
  • Author(s): Kobayashi, T.
  • Organizer: Gordon Reseach Conference on Glycolipid and Sphingolipid Biology
  • Place of Presentation: Lucca, Italy
• [Presentation] 細胞質分裂におけるスフィンゴミエリンドメインの役割 (2012)
  • Author(s): 小林俊秀
  • Organizer: 第８５回日本生化学会
  • Place of Presentation: 福岡
  • Invited
• [Presentation] エンドソーム特異的脂質の構造、代謝、機能 (2012)
  • Author(s): 小林俊秀
  • Organizer: 第５４回日本脂質生化学会
  • Place of Presentation: 福岡
  • Invited
• [Presentation] A role for sphingomyelin-rich lipid domains in the accumulation of phosphatidylinositol-4,5-bisphosphate to the cleavage furrow during cytokinesis (2012)
  • Author(s): Toshihide Kobayashi
  • Organizer: Lipid-Protein Interactions in Membranes: Implications for Health and Disease
  • Place of Presentation: Hyderabad
  • Invited
• [Presentation] Imaging lipids and lipid domains (2011)
  • Author(s): Kobayashi T.
  • Organizer: International ERATO Symposium on Lipid Structures in and around Proteins
  • Place of Presentation: 阪急エキスポパーク(大阪)(招待講演)
  • Year and Date: 2011-11-12
• [Presentation] Imaging lipid domains (2011)
  • Author(s): Kobayashi T.
  • Organizer: 30^<th> Naito conference, "Membrane Dynamics and Lipid Biology [II]"
  • Place of Presentation: シャトレーゼガトーキングダム札幌(札幌)(招待講演)
  • Year and Date: 2011-06-29
• [Presentation] Imaging lipids and lipid domains (2011)
  • Author(s): Kobayashi, T.
  • Organizer: International ERATO Symposium on Lipid Structures in and around Proteins
  • Place of Presentation: Osaka
• [Presentation] 脂質のダイナミズムを見る-脂質・蛋白質相互作用の解析 (2011)
  • Author(s): 小林俊秀
  • Organizer: 第34回日本神経科学大会
  • Place of Presentation: 横浜
• [Presentation] Imaging lipid domains Naito conference, (2011)
  • Author(s): Kobayashi, T., Abe, M., Ishitsuka, R., Kishimoto, T., Makino, A., Hullin-Matsuda, F.,and M. Murate
  • Organizer: 30^(th)Naito conference "Membrane Dynamics and Lipid Biology [II]"
  • Place of Presentation: Sapporo
• [Presentation] Visualization of lipids and lipid domains (2011)
  • Author(s): 小林俊秀、阿部充宏、岸本拓磨、牧野麻美、Hullin-Matsuda Francoise、石塚玲子、村手源英
  • Organizer: 第53回日本脂質生化学会
  • Place of Presentation: 東京
• [Presentation] Imaging of membrane heterogeneity (2010)
  • Author(s): Kobayashi, T.
  • Organizer: 4^<th> ISN Special Conference on Membrane Domains in CNS physiology and pathology
  • Place of Presentation: Erice, Italy
  • Year and Date: 2010-05-22
• [Presentation] 脂質ラフトのイメージング (2010)
  • Author(s): 小林俊秀
  • Organizer: 第3回セラミド研究会学術集会
  • Place of Presentation: 東京
• [Presentation] Tool box for imaging lipids and lipid domains (I) (II) (2010)
  • Author(s): Kobayashi, T.
  • Organizer: ISN International School: Lipidomics in the nervous system cells
  • Place of Presentation: Catania, Italy,
• [Remarks]
  • URL: http://www.riken.jp/lbl/mainpagelbl.html
• [Remarks]
  • URL: http://www.riken.jp/lbl/mainpagelbl.html
• [Remarks]
  • URL: http://www.riken.jp/lbl/mainpagelbl.html
• [Patent(Industrial Property Rights)] 新規なタンパク質およびその利用 (2010)
  • Inventor(s): 小林俊秀
  • Industrial Property Rights Holder: 理化学研究所
  • Filing Date: 2010-05-14