Dynamic analysis of transport vesicle formation using artificial planar lipid bilayer

• Project/Area Number: 19687008
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Functional biochemistry
• Research Institution: The University of Tokyo
• Principal Investigator: SATO Ken The University of Tokyo, 大学院・総合文化研究科, 准教授 (00303602)
• Project Period (FY): 2007 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥25,480,000 (Direct Cost: ¥19,600,000、Indirect Cost: ¥5,880,000); Fiscal Year 2009: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2008: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2007: ¥12,480,000 (Direct Cost: ¥9,600,000、Indirect Cost: ¥2,880,000)
• Keywords: 小胞輸送 / 輸送小胞 / 小胞体 / 低分子量GTPase / 出芽酵母 / 1分子計測 / プロテオリポソーム / GTPase

Research Abstract

We investigated the dynamics of fluorescently labeled cargo proteins during COPII vesicle formation using single-molecule microscopy combined with an artificial planar lipid bilayer. We show that continuous GTPase cycles of Sar1 facilitate cargo concentration into COPII vesicle buds. We propose that the minimal set of COPII components is required to concentrate cargo molecules.

Research Products

• [Journal Article] Structural basis for the Rho- and phosphoinositide-dependent localization of the exocyst subunit Sec3. (2010)
  • Author(s): Yamashita, M., Kurokawa, K., Sato, Y., Yamagata, A., Mimura, H., Yoshikawa, A. Sato, K., Nakano, A., Fukai, S.
  • Journal Title: Nature Struct. &Mol. Biol. 17 Pages: 180-186
  • Peer Reviewed
• [Journal Article] Structural basis for the Rho- and phosphoinositide-dependent localization of the exocyst subunit Sec3 (2010)
  • Author(s): Yamashita, M., et al.
  • Journal Title: Nature Struct & Mel.Biol. 17 Pages: 180-186
  • Peer Reviewed
• [Journal Article] Visualization of cargo concentration by COPII minimal machinery in a planar lipid membrane. (2009)
  • Author(s): Tabata, K.V., Sato, K., Ide, T., Nishizaka, T., Nakano, A., Noji, H.
  • Journal Title: EMBO J. 28 Pages: 3279-3289
  • Peer Reviewed
• [Journal Article] O-Mannosylation is required for the solubilization of heterologously expressed human b-amyloid precursor protein in Saccharomyces cerevisiae. (2009)
  • Author(s): Murakami-Sekimata, A., Sato, K., Sato, K., Takashima, A., Nakano, A.
  • Journal Title: Genes Cells 14 Pages: 205-215
  • Peer Reviewed
• [Journal Article] Visualization of cargo concentration by COPII minimal machinery in a planar lipid membrane (2009)
  • Author(s): Tabata, K.V., et al.
  • Journal Title: EMBO J. 28 Pages: 3279-3289
  • Peer Reviewed
• [Journal Article] O-Mannosylation is required for the solubilization of heterologously expressed human b-amyloid precursor protein in S. cerevisiae (2009)
  • Author(s): Murakami-Sekimata, A. et.al.
  • Journal Title: Genes Cells 14 Pages: 205-215
  • Peer Reviewed
• [Journal Article] Smy2p participates in COPII vesicle formation through the interaction with Sec23p/Sec24p subcomplex. (2008)
  • Author(s): Higashio, H., Sato, K., Nakano, A.
  • Journal Title: Traffic 9 Pages: 79-93
  • Peer Reviewed
• [Journal Article] Smy2p participates in COPII vesicle formation through the interaction with Sec23p/Sec24p subcomplex (2008)
  • Author(s): Hironori Higashio, et. al.
  • Journal Title: Traffic 9 Pages: 79-93
  • Peer Reviewed
• [Journal Article] Mechanisms of COPII vesicle formation and protein sorting. (2007)
  • Author(s): Sato, K., Nakano, A.
  • Journal Title: FEBS Lett. 581 Pages: 2076-2082
  • Peer Reviewed
• [Journal Article] VPS9a, the common activator for two distinct types of Rab5 GTPases, is essential for the development of Arabidopsis thaliana. (2007)
  • Author(s): Goh, T., Uchida, W., Arakawa, S., Ito, E., Dainobu, T., Ebine, K., Takeuchi, M., Sato, K., Ueda, T., Nakano, A.
  • Journal Title: Plant Cell 19 Pages: 3504-3515
  • Peer Reviewed
• [Journal Article] Mechanisms of COPII vesicle formation and protein sorting (2007)
  • Author(s): Ken Sato, et. al.
  • Journal Title: FEBS Lett. 581 Pages: 2076-2082
  • Peer Reviewed
• [Journal Article] VPS9a, the common activator for two distinct types of Rab5 GTPases, is essential for the development of Arabidopsis thaliana (2007)
  • Author(s): Tatsuaki Goh, et. al.
  • Journal Title: Plant Cell 19 Pages: 3504-3515
  • Peer Reviewed
• [Presentation] 小胞体における輸送小胞形成の分子メカニズムの解析 (2009)
  • Author(s): 佐藤健
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2009-10-21
• [Presentation] Dynamics of COPII-mediated cargo concentration observed in a planar lipid membrane (2008)
  • Author(s): Ken Sato
  • Organizer: 第31回日本分子生物学会・第81回日本生化学会合同大会
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2008-12-12
• [Presentation] Sed4pによるSar1pの制御機構の解析 (2008)
  • Author(s): 小寺千絵、中野明彦、佐藤健
  • Organizer: 第31回日本分子生物学会・第81回日本生化学会合同大会
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2008-12-12
• [Presentation] Dynamics of COPII-mediated cargo concentration observed in a planar lipid membrane (2008)
  • Author(s): Ken Sato
  • Organizer: 第31回日本分子生物学会・第81回日本生化学会合同大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2008-12-12
• [Presentation] Sed4pによるSar1pの制御機構の解析 (2008)
  • Author(s): 小寺千絵, ほか
  • Organizer: 第31回日本分子生物学会・第81回日本生化学会合同大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2008-12-12
• [Presentation] Cargo concentration and COPII vesicle formation observed in reconstituted planar model membrane system (2007)
  • Author(s): Ken Sato
  • Organizer: 第30回日本分子生物学会・第80回日本生化学会合同大会
  • Year and Date: 2007-12-13
• [Presentation] Cargo concentration and COPII vesicle formation observed in reconstituted planar model membrane system (2007)
  • Author(s): Ken Sato
  • Organizer: 第30回日本分子生物学会・第80回日本生化学会大会合同大会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-13
• [Book] 細胞工学(Sar1が制御する輸送小胞形成の分子メカニズム) (2009)
  • Author(s): 佐藤健
  • Publisher: 秀潤社
• [Book] 細胞工学「Sar1が制御する輸送小胞形成の分子メカニズム」 (2009)
  • Author(s): 佐藤健
  • Total Pages: 5
  • Publisher: 秀潤社
• [Book] The Golgi Apparatus-COPII (2008)
  • Author(s): Ken Sato, Akihiko Nakano
  • Publisher: Springer
• [Book] 蛋白質 核酸 酵素(小胞体における積み荷蛋白質の選別とCOPII小胞京成のメカニズム) (2008)
  • Author(s): 佐藤健
  • Publisher: 共立出版
• [Book] The Golgi Apparatus (2008)
  • Author(s): Sato, K. et.al.
  • Publisher: Springer Wien New York
• [Book] 小胞体における積み荷蛋白質の選別とCOPII小胞形成のメカニズム (2008)
  • Author(s): 佐藤健
  • Publisher: 共立出版