Electron microscopic analysis of lipid clusters and membrane proteins in plasma membrane.

• Project/Area Number: 23590251
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General anatomy (including Histology/Embryology)
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: 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)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2012: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2011: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 脂質ドメイン / フリーズフラクチャー / 脂質結合タンパク質 / 血小板 / マイクロパーティクル / スフィンゴミエリン / ホスファチジルセリン / 細胞膜 / 電子顕微鏡 / 脂質ラフト / 透過電子顕微鏡 / リポソーム / ホスファチジルイノシトール-4,5-2リン酸 / 免疫電子顕微鏡法 / フリーズフラクチャー法 / カベオラ / リン脂質 / コレステロール

Research Abstract

We examined transbilayer and lateral lipid distribution in plasma membrane using recently developed peptide/proteins that selectively bind specific lipids or lipid clusters. The liposome experiments demonstrated that the lipids were effectively detected by the probes even when the membrane concentration was as low as 1 mol%, and the probes bound similarly to the model membranes that mimic outer or inner leaflet lipid environment.
Next, we examined the distribution of sphingomyelin and phosphatidylserine in resting and thrombin-stimulated platelets as well as platelet microparticles that are highly procoagulant. Our results indicate that the scrambled membranes are selectively released in the form of microparticles in thrombin-activated platelets and suggest the idea that the exposure of phosphatidylserine in inner leaflet to the outer triggers the vesiculation of platelets.

Research Products

• [Journal Article] Binding of a pleurotolysin ortholog from Pleurotus eryngii to sphingomyelin and cholesterol-rich membrane domains. (2013)
  • Author(s): Bhat H B, Kishimoto T, Abe M, Makino A, Inaba T, Murate M, Dohmae N, Kurahashi A, Nishibori K, Fujimori F, Greimel P, Ishitsuka R, Kobayashi T.
  • Journal Title: Journal of Lipid Research Volume: 54 Issue: 10 Pages: 2933-2943
  • DOI: 10.1194/jlr.d041731
  • Peer Reviewed
• [Journal Article] Phosphatidylglucoside: Its structure, thermal behavior, and domain formation in plasma membranes. (2012)
  • Author(s): Hiroshi Takahashi, Tomohiro Hayakawa, Motohide Murate, Peter Greimel, Yasuko Nagatsuka, Toshihide Kobayashi, Yoshio Hirabayashi.
  • Journal Title: Chemistry and Physics of Lipids Volume: 165 Issue: 2 Pages: 197-206
  • DOI: 10.1016/j.chemphyslip.2011.12.010
  • Peer Reviewed
• [Journal Article] Evaluation of the influence of ionization states and spacers in the thermotropic phase behaviour of amino acid-based cationic lipids and the transfection efficiency of their assemblies (2012)
  • Author(s): Sarker SR, Arai S, Murate M, Takahashi H, Takata M, Kobayashi T, Takeoka S
  • Journal Title: Int. J. Pharma Volume: 422 Pages: 364-373
  • Peer Reviewed
• [Journal Article] Evaluation of the influence of ionization states and spacers in the thermotropic phase behaviour of amino acid-based cationic lipids and the transfection efficiency of their assemblies. (2012)
  • Author(s): Satya Ranjan Sarker, Satoshi Arai, Motohide Murate, Hiroshi Takahashi, Masaki Takata, Toshihide Kobayashi, Shinji Takeoka.
  • Journal Title: International Journal of Pharmacology Volume: 42 Issue: 1-2 Pages: 364-373
  • DOI: 10.1016/j.ijpharm.2011.10.044
  • Peer Reviewed
• [Presentation] Transbilayer and lateral lipid distributions in nano scale. (2014)
  • Author(s): Murate M, Abe M, Kasahara K, Iwabuchi K, Umeda M, Kobayashi T.
  • Organizer: EMBO Workshop "Cellular imaging of lipids"
  • Place of Presentation: Vico Equense, Italy
• [Presentation] Transbilayer lipid distribution on the plasma membrane monitored by SDS-freeze fracture replica method (2013)
  • Author(s): Murate M
  • Organizer: Scientific Symposium on Applications of Advanced Microscopy Technique in Materials and Life Sciences
  • Place of Presentation: Ljubuljana, Slovenia
  • Year and Date: 2013-09-20
• [Presentation] Membrane deformation by phospholipase C beta 1 (2013)
  • Author(s): 岸本拓磨、稲葉岳彦、田島拓也、阿部充宏、村手源英、牧野麻美、石塚玲子、池田康夫、武岡真司、小林俊秀
  • Organizer: 理研シンポジウム脂質・タンパク質相互作用:分子から細胞へ
  • Place of Presentation: 和光
  • Year and Date: 2013-05-24
• [Presentation] Cardiolipin polymorphism : Molecular mechanism of inhibition (2013)
  • Author(s): Greimel P, Murate M, Hullin-Matsuda F, Iwamoto K, Ito K, Takata M, Kobayashi M, Takahashi H, Kobayashi T
  • Organizer: 54th International Conference on the Biosciences of Lipids
  • Place of Presentation: Bari, Italy
• [Presentation] Cardiolipin polymorphism : Molecular mechanism of inhibition by 10-N-nonyl acridine orange. Cardiolipin as key lipid of mitochondria in health and disease (2013)
  • Author(s): Greimel P, Murate M, Hullin-Matsuda F, Iwamoto K, Ito K, Takata M, Kobayashi M, Takahashi H, Kobayashi T
  • Organizer: 54th International Conference on the Biosciences of Lipids
  • Place of Presentation: Bari, Italy
• [Presentation] Free cholesterol loading induces the degradation of perilipin 2 and rab18-dependent 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
• [Presentation] Transformation of phosphatidylethanolamine- and phosphatidylserine-containing membranes by phospholipase C beta 1 (2013)
  • Author(s): Tajima T, Inaba T, Kishimoto T, Abe M, Murate M, Makino A, Sakai S, Hullin-Matsuda F, Ishitsuka R, Ikeda Y, Takeoka S, Kobayashi T
  • Organizer: 14th International Membrane Research Forum
  • Place of Presentation: Kyoto
• [Presentation] Revisiting the interaction of 10 N-nonyl acridine orange with cardiolipin (2013)
  • Author(s): Greimel P, Murate M, Takahashi H, Kobayashi T
  • Organizer: 14th International Membrane Research Forum
  • Place of Presentation: Kyoto
• [Presentation] Binding of pleurotolysin ortholog from Pleuroyus eryngii to sphingomyelin and cholesterol-rich membrane domains (2013)
  • Author(s): Bhat HB, Kishimoto T, Abe M, Makino A, Inaba T, Murate M, Dohmae N, Kurahashi A, Nishibori K, Fujimori F, Ishitsuka R, Kobayashi T
  • Organizer: 14th International Membrane Research Forum
  • Place of Presentation: Kyoto
• [Presentation] Cardiolipin polymorphism: Molecular mechanism of inhibition. (2013)
  • Author(s): Greimel P, Murate M, Hullin-Matsuda F, Iwamoto K, Ito K, Takata M, Kobayashi M, Takahashi H, Kobayashi T.
  • Organizer: 54th International Conference on the Biosciences of Lipids
  • Place of Presentation: Bari, Italy
• [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: 神奈川県横浜市
• [Presentation] Binding of pleurotolysin ortholog from Pleuroyus eryngii to sphingomyelin and cholesterol-rich membrane domains. (2013)
  • Author(s): Balakrishna B.H., Kishimoto T., Abe M., Makino A., Inaba T., Murate M., Dohmae N., Kurahashi A., Nishibori K., Fujimori F., Ishitsuka R. and Kobayashi T.
  • Organizer: 14th International Membrane Research Forum.
  • Place of Presentation: 京都府京都市
• [Presentation] Revisiting the interaction of 10 N-nonyl acridine orange with cardiolipin. (2013)
  • Author(s): Peter Greimel , Motohide Murate, Hiroshi Takahashi and Toshihide Kobayashi.
  • Organizer: 14th International Membrane Research Forum.
  • Place of Presentation: 京都府京都市
• [Presentation] Transformation of phosphatidylethanolamine- and phosphatidylserine-containing membranes by phospholipase C beta 1. (2013)
  • Author(s): Tajima T., Inaba T., Kishimoto T., Abe M., Murate M., Makino A., Sakai S., Hullin-Matsuda F., Ishitsuka R., Ikeda Y., Takeoka S. and Kobayashi T.
  • Organizer: 14th International Membrane Research Forum.
  • Place of Presentation: 京都府京都市
• [Presentation] Phospholipase C-beta 1b changes the shape of phosphatidylethaonolamine- containing membranes (2012)
  • Author(s): Tajima T, Inaba T, Kishimoto T, Abe M, Murate M, Sakai S, Ishitsuka R, Ikeda Y, Takeoka S, Kobayashi T
  • Organizer: RIKEN Nano Science Joint Laboratory Forum
  • Place of Presentation: Wako
  • Year and Date: 2012-11-29
• [Presentation] Phospholipase C-beta 1b changes the shape of phosphatidylethaonolamine- containing membranes (2012)
  • Author(s): Tajima T, Inaba T, Kishimoto T, Abe M, Murate M, Sakai S, Ishitsuka R, Ikeda Y, Takeoka S, Kobayashi T
  • Organizer: Lipid-Protein Interactions in Membranes : Implications for Health and Disease
  • Place of Presentation: Hyderabad, India
• [Presentation] Revisiting plasma membrane lipid asymmetry (2012)
  • Author(s): Murate M, Abe M, Umeda M, Kobayashi T
  • Organizer: 53rd International Conference on the Bioscience of Lipids
  • Place of Presentation: Banff, Canada
• [Presentation] Molecular mechanism of the inhibition of cardiolipin polymorphism by 10-N-Nonyl Acridine Orange (2012)
  • Author(s): Greimel P, Murate M, Hullin-Matsuda F, Nabetani T, Iwamoto K, Ito K, Takata M, Kobayashi M, Takahashi H, Kobayashi T
  • Organizer: 53rd International Conference on the Bioscience of Lipids
  • Place of Presentation: Banff, Canada
• [Presentation] Phospholipase C-beta 1b changes the shape of phosphatidylethaonolamine-containing membranes. (2012)
  • Author(s): Tajima T., Inaba T., Kishimoto T., Abe M., Murate M., Sakai S., Ishitsuka R., Ikeda Y., Takeoka S. and Kobayashi T.
  • Organizer: Lipid-Protein Interactions in Membranes: Implications for Health and Disease.
  • Place of Presentation: Hyderabad, India
• [Presentation] Molecular mechanism of the inhibition of cardiolipin polymorphism by 10-N-Nonyl Acridine Orange. (2012)
  • Author(s): Greimel P., Murate M., Hullin-Matsuda F., Nabetani T., Iwamoto K., Ito K., Takata M., Kobayashi M., Takahashi H. and Kobayashi T.
  • Organizer: 53rd International Conference on the Bioscience of Lipids.
  • Place of Presentation: Banff, Canada
• [Presentation] Revisiting plasma membrane lipid asymmetry. (2012)
  • Author(s): Motohide Murate, Mitsuhiro Abe, Masato Umeda and Toshihide Kobayashi.
  • Organizer: 53rd International Conference on the Bioscience of Lipids.
  • Place of Presentation: Banff, Canada
• [Presentation] Imaging lipids and lipid domains (2011)
  • Author(s): Greimel P, Murate M, Hullin-Matsuda F, Nabetani T, Iwamoto K, Ito K, Takata M, Takahashi H, Kobayashi T
  • Organizer: International Symposium on Lipid Structures in and around Proteins
  • Place of Presentation: Osaka
• [Presentation] 新規糖リン脂質ホスファチジルグルコシドの転移温度・エンタルピー値について (2011)
  • Author(s): 高橋浩、早川智広、村手源英、Greimel P、長塚靖子、小林俊秀、平林義雄
  • Organizer: 第63回コロイドおよび界面化学討論会
  • Place of Presentation: 京都
• [Presentation] Endplasmic reticulm cholesterol controls both ApoB serection and the formation of lipid droplets (2011)
  • Author(s): 牧野麻美、Hullin-Matsuda F、村手源英、阿部充宏、Vandenbroucke I、Lagarde M、小林俊秀
  • Organizer: 第30回内藤コンファレンス「生体膜ダイナミクスと脂質生物学[II]:脂質ドメイン、脂肪滴、疾患」
  • Place of Presentation: 札幌
• [Presentation] Imaging lipid domains (2011)
  • Author(s): 小林俊秀、阿部充宏、石塚玲子、岸本拓磨、牧野麻美、Hullin-Matsuda F、村手源英
  • Organizer: 第30回内藤コンファレンス「生体膜ダイナミクスと脂質生物学[II]:脂質ドメイン、脂肪滴、疾患」
  • Place of Presentation: 札幌
• [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 F、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 lipid domains. (2011)
  • Author(s): 小林俊秀, 阿部充宏, 石塚玲子, 岸本拓磨, 牧野麻美, Hullin-Matsuda Francoise, 村手源英.
  • Organizer: 第30回内藤コンファレンス「生体膜ダイナミクスと脂質生物学[II]：脂質ドメイン、脂肪滴、疾患」(招待講演)
  • Place of Presentation: シャトレーゼガトーキングダムサッポロ
• [Presentation] Endplasmic reticulm cholesterol controls both ApoB serection and the formation of lipid droplets. (2011)
  • Author(s): 牧野麻美, Hullin-Matsuda Francoise, 村手源英, 阿部充宏, Vandenbroucke Isabelle, Lagarde Michel, 小林俊秀.
  • Organizer: 第30回内藤コンファレンス「生体膜ダイナミクスと脂質生物学［II］：脂質ドメイン、脂肪滴、疾患」
  • Place of Presentation: シャトレーゼガトーキングダムサッポロ
• [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(招待講演)
  • Place of Presentation: 東京ガーデンパレス
• [Presentation] Transbilayer lipid distribution on the plasma membrane monitored by SDS-freeze fracture replica method.
  • Author(s): Murate M.
  • Organizer: Scientific Symposium on Applications of Advanced Microscopy Technique in Materials and Life Sciences
  • Place of Presentation: Ljubuljana, Slovenia
  • Invited
• [Presentation] Membrane deformation by phospholipase C beta 1.
  • Author(s): 岸本拓磨、稲葉岳彦、田島拓也、阿部充宏、村手源英、牧野麻美、石塚玲子、Ikeda Yasuko、Takeoka Shinji、小林俊秀
  • Organizer: 理研シンポジウム 脂質・タンパク質相互作用：分子から細胞へ
  • Place of Presentation: 埼玉県和光市
• [Remarks]
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• [Remarks] Welcome to Lipid Biology Laboratory!
  • URL: http://www.riken.jp/lbl/mainpagelbl.html
• [Remarks] 小林脂質生物学研究室
  • URL: http://www.asi.riken.jp/jp/laboratories/chieflabs/lipid/
• [Remarks] 小林脂質生物学研究室
  • URL: http://www.riken.jp/lbl/mainpagelbl.html
• [Remarks]
  • URL: http://www.riken.jp/lbl/mainpagelbl.html
• [Remarks]
  • URL: http://www.asi.riken.jp/jp/laboratories/chieflabs/lipid/index.html