The structural analysis of the sulfonylurea receptor by single particle analysis

• Project/Area Number: 19790034
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Physical pharmacy
• Research Institution: Kyoto University
• Principal Investigator: KIMURA Yasuhisa Kyoto University, 大学院・農学研究科, 助教 (10415143)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥3,780,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥480,000); Fiscal Year 2008: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2007: ¥1,700,000 (Direct Cost: ¥1,700,000)
• Keywords: スルフォニル尿素受容体 / 単粒子解析 / ATP感受性カリウムチャネル / KATP / SUR1 / Kir6.2 / ABC transporter / single particle analysis

Research Abstract

本研究ではヒト培養細胞を用いた哺乳類由来の膜タンパク質複合体につき、発現精製に必要な基盤技術を開発した。またこれらの技術を用いてスルフォニル尿素剤受容体を発現精製し、単粒子解析により基準となる構造の決定を行った。また、タンパク質工学的手法によりヒトMDR1のリンカードメインが持つ機能を明らかにした。さらにヒトABCA1のシステイン結合が機能に必須であることを明らかにした。

Research Products

• [Journal Article] Formation of two intramolecular disulfide bonds is necessary for apoA-I-dependent cholesterol efflux mediated by ABCA1 (2009)
  • Author(s): Hozoji, M., Kimura, Y., Kioka, N., Ueda, K.
  • Journal Title: J. Biol. Chem. 284 Pages: 11293-11300
  • Peer Reviewed
• [Journal Article] Sodium taurocholate-dependent lipid efflux by ABCA1-Effects of W590S mutation on lipid translocation and apoA-I dissociation (2009)
  • Author(s): Nagao, K. Zhao, Y., Takahashi, K., Kimura, Y., Ueda, K.
  • Journal Title: J. Lipid Res. 50 Pages: 1165-1172
  • Peer Reviewed
• [Journal Article] Functional role of the linker region in purified human P-glycoprotein (2009)
  • Author(s): Sato, T., Kodan, A., Kimura, Y., Ueda, K., Nakatsu, T. and Kato, H
  • Journal Title: FEBS J. (in press)
  • Peer Reviewed
• [Journal Article] Formation of two intramolecular disulfide bonds is necessary for apoA-I-dependent cholesterol efflux mediated by ABCAl. (2009)
  • Author(s): Hozoji, M., Kimura, Y., Kioka, N. and Ueda, K.
  • Journal Title: J. Biol. Chem. (in press)
  • Peer Reviewed
• [Journal Article] Sodium taurocholate-dependent lipid efflux by ABCAl-Effects of W590S mutation on lipid translocation and apoA-I dissociation. (2009)
  • Author(s): Nagao, K. Zhao, Y., Takahashi, K., Kimura, Y., and Ueda, K.
  • Journal Title: J. Lipid Res (in press)
  • Peer Reviewed
• [Journal Article] Mechanism of multi-drug recognition by MDR1/ABCB1 (2007)
  • Author(s): Kimura, Y., Morita, S.-y., Matsuo, M., Ueda, K.
  • Journal Title: Cancer Sci. Pages: 1303-1310
  • NAID: 10019998059
  • Peer Reviewed
• [Journal Article] Functional role of the linker region in purified human P-glycoprotein
  • Author(s): Sato, T., Kodan, A., Kimura, Y., Ueda, K., Nakatsu, T., Kato, H.
  • Journal Title: FEBS J. (in press)
  • Peer Reviewed
• [Presentation] Large scale production and purification of KATP channel in mammalian cells (2009)
  • Author(s): Y. Kimura, H. Kato, K. Ueda
  • Organizer: Takeda Science Foundation Symposium on PharmaSciences
  • Place of Presentation: Tokyo
  • Year and Date: 2009-05-24
• [Presentation] ヒトABCG1の大量培養系の構築・安定化条件の探索 (2009)
  • Author(s): 平山裕士, 木村泰久, 松尾道憲, 木岡紀幸, 植田和光
  • Organizer: 農芸化学会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-28
• [Presentation] ヒト培養細胞を用いたATP感受性カリウムチャネル複合体の大量生産、高純度精製 (2009)
  • Author(s): 木村泰久, 平山裕士, 松尾道憲, 木岡紀幸, 加藤博章, 植田和光
  • Organizer: 農芸化学会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-28
• [Presentation] ヒト培養細胞を用いたATP感受性カリウムチャネル復合体の大量生産、高純度精製 (2009)
  • Author(s): 木村泰久、平山裕士、加藤博章、植田和光
  • Organizer: 農芸化学会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-28
• [Presentation] ヒト培養細胞を用いたヒトABCトランスポーターの大量発現系の構築および精製条件スクリーニング手法の開発 (2008)
  • Author(s): 木村泰久, 平山裕士, 浅見成美, 木岡紀幸, 松尾道憲, 加藤博章, 植田和光
  • Organizer: 分子生物学会
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-15
• [Presentation] Mechanism of multidrug recognition by MDR1 - cholesterol fill-in model (2008)
  • Author(s): 木村泰久, 松尾道憲, 木岡紀幸, 植田和光
  • Organizer: FEBS-ABC2008
  • Place of Presentation: Innsbruck, Austria