Visualization and analysis of receptor oligomerization in living cells by using novel fluorescence labeling method

• Project/Area Number: 21390007
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Physical pharmacy
• Research Institution: Kyoto University
• Principal Investigator: MATSUZAKI Katsumi 京都大学, 大学院・薬学研究科, 教授 (00201773)
• Co-Investigator(Kenkyū-buntansha): YANO Yoshiaki 京都大学, 大学院・薬学研究科, 助教 (60402799)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥18,850,000 (Direct Cost: ¥14,500,000、Indirect Cost: ¥4,350,000); Fiscal Year 2011: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000); Fiscal Year 2010: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000); Fiscal Year 2009: ¥13,130,000 (Direct Cost: ¥10,100,000、Indirect Cost: ¥3,030,000)
• Keywords: 蛍光イメージング / GPCR / オリゴマー化 / βアドレナリン受容体 / 蛍光励起エネルギー移動 / コンフォメーション変化 / β2アドレナリン受容体 / グルタミン酸受容体

Research Abstract

An accurate method to analyze oligomeric state of membrane proteins in living cells by using novel fluorescence labeling method(coiled-coil labeling method) has been established. The method enables analysis of association number of target proteins, which has been difficult by using conventional methods.

Research Products

• [Journal Article] Coiled-coil tag-probe labeling method for live-cell imaging of membrane receptors (2012)
  • Author(s): Yoshiaki Yano, Kenichi Kawano, Kaoru Omae, and Katsumi Matsuzaki
  • Journal Title: Methods Enzymol Volume: 504 Pages: 355-370
  • DOI: 10.1016/b978-0-12-391857-4.00018-5
• [Journal Article] 新規蛍光ラベル法による膜受容体の内在化の可視化 (2010)
  • Author(s): 松崎勝巳, 矢野義明
  • Journal Title: 生化学 Volume: 82 Pages: 494-497
  • NAID: 10026899941
  • URL: http://www.jbsoc.or.jp/event/magazine/index.html
• [Journal Article] Design of a soluble transmembrane helix for measurements of water-membrane partitioning (2010)
  • Author(s): Yoshiaki Yano, Norie Shimai, Katsumi Matsuzaki
  • Journal Title: J.Phys.Chem.B. 114 Pages: 1925-1931
  • Peer Reviewed
• [Journal Article] Tag-probe fluorescent labeling methods for live-cell imaging of membrane proteins Biochim (2009)
  • Author(s): Yoshiaki Yano and Katsumi Matsuzaki
  • Journal Title: Biophys. Acta Volume: 1788 Issue: 10 Pages: 2124-2131
  • DOI: 10.1016/j.bbamem.2009.07.017
• [Journal Article] Peptide-lipid huge toroidal pore : a new antimicrobial mechanism mediated by a lactococcal bacteriocin, lacticin Q (2009)
  • Author(s): Fuminori Yoneyama, Yuichi Imura, Kanako Ohno, Takeshi Zendo, Jiro Nakayama, Katsumi Matsuzaki, Kenji Sonomoto
  • Journal Title: Antimicrob.Agents Chemother 53 Pages: 3211-3217
  • Peer Reviewed
• [Journal Article] Ganglioside-induced formation of amyloid fibrils by hIAPP in lipid rafts (2009)
  • Author(s): Masaki Wakabayashi, Katsumi Matsuzaki
  • Journal Title: FEBS Lett. 583 Pages: 2854-2858
  • Peer Reviewed
• [Journal Article] Tag-probe fluorescent labeling methods for live-cell imaging of membrane proteins (2009)
  • Author(s): Yoshiaki Yano, Katsumi Matsuzaki
  • Journal Title: Biochim.Biophys.Acta 1788 Pages: 2124-2131
  • Peer Reviewed
• [Journal Article] (2009)
  • Author(s): 松崎勝巳, 矢野義明
  • Journal Title: 薬学分析化学の最前線(じほう) Pages: 170-171
• [Presentation] Internalization and oligomerization of membrane receptors in living cells as detected by novel coiled-coil fluorescence labeling methods (2011)
  • Author(s): Katsumi Matsuzaki, Kaoru Omae, Yoshiaki Yano
  • Organizer: International Peptide Symposium
  • Place of Presentation: Kyoto International Conference Center(京都府)
  • Year and Date: 2011-12-06
• [Presentation] Detection of oligomerization and internalization of membrane receptors in living cells by novel coiled-coil fluorescence labeling method (2011)
  • Author(s): Yano Y, Kawano K, Omae K, Matsuzaki K
  • Organizer: AIMECS11
  • Place of Presentation: 京王プラザホテル(東京都)
  • Year and Date: 2011-11-29
• [Presentation] Behaviors of receptors as analyzed by novel fluorescence labeling method for membrane proteins (2011)
  • Author(s): 松崎勝巳
  • Organizer: 第49回日本生物物理学会シンポジウム
  • Place of Presentation: 兵庫県立大学(兵庫県)
  • Year and Date: 2011-09-16
• [Presentation] Behaviors of receptors as analyzed by novel fluorescence labeling method for membrane proteins (2011)
  • Author(s): 松崎勝巳
  • Organizer: 第49回日本生物物理学会シンポジウム
  • Place of Presentation: 兵庫県立大学(兵庫県)(招待講演)
  • Year and Date: 2011-09-16
• [Presentation] Family A GPCRの受容体活性に会合体形成は本当に必要か (2011)
  • Author(s): 河野健一、大前薫、矢野義明、松崎勝巳
  • Organizer: 第9回次世代を担う若手のためのフィジカル・ファーマフォーラム
  • Place of Presentation: ホテル箱根アカデミー(神奈川県)
  • Year and Date: 2011-09-12
• [Presentation] Internalization and oligomerization of membrane receptors in living cells as detected by novel coiled-coil fluorescence labeling methods (2010)
  • Author(s): Katsumi Matsuzaki, Kaoru Omae, Yoshiaki Yano
  • Organizer: 5^<th> International Peptide Symposium
  • Place of Presentation: Kyoto International Conference Center(京都府)
  • Year and Date: 2010-12-06
• [Presentation] コイルドコイル蛍光ラベル法:次世代の膜受容体可視化ツール (2010)
  • Author(s): 矢野義明、大前薫、竹田有希、松崎勝巳
  • Organizer: 第32回生体膜と薬物の相互作用シンポジウム
  • Place of Presentation: 富山国際会議場(富山県)
  • Year and Date: 2010-11-29
• [Presentation] Oligomerization of membrane receptors (2010)
  • Author(s): Yoshiaki Yano, Kaoru Omae, and Katsumi Matsuzaki
  • Organizer: FRET analysis using coiled-coil tag-probe labeling and spectral imaging Biophysical Society 54th annual meeting Aan Francisco
  • Place of Presentation: USA
  • Year and Date: 2010-10-23
• [Presentation] 新規蛍光標識法による膜受容体の可視化解析とリガンドスクリーニング (2010)
  • Author(s): 松崎勝巳
  • Organizer: 第28回物性物理化学研究会
  • Place of Presentation: 京都大学(京都府)
  • Year and Date: 2010-06-04
• [Presentation] 新規小分子蛍光ラベル法による生細胞でのGPCRオリゴマーの検出 (2010)
  • Author(s): 大前薫, 矢野義明, 松崎勝巳
  • Organizer: 日本薬学会 第130年会
  • Place of Presentation: 岡山
  • Year and Date: 2010-03-29
• [Presentation] 新規小分子蛍光ラベル法による生細胞でのGPCRオリゴマーの検出 (2010)
  • Author(s): 大前薫、矢野義明、松崎勝巳
  • Organizer: 日本薬学会第130年会岡山大学
  • Place of Presentation: 岡山県
  • Year and Date: 2010-03-28
• [Presentation] 新規蛍光ラベル法を用いた生細胞膜での受容体オリゴマー解析法の開発 (2010)
  • Author(s): 矢野義明
  • Organizer: 第8回次世代を担う若手のためのフィジカル・ファーマフォーラム
  • Place of Presentation: 岡山 (岡山大学)
  • Year and Date: 2010-03-26
• [Presentation] OLIGOMERIZATION OF MEMBRANE RECEPTORS : FRET ANAL YSIS USING COILED-COIL TAG-PROBE LABELING AND SPECTRAL IMAGING (2010)
  • Author(s): Yoshiaki Yano, Kaoru Omae, Katsumi Matsuzaki
  • Organizer: Biophysical Society 54th Annual Meeting
  • Place of Presentation: San Francisco (Moscone Cantor), USA
  • Year and Date: 2010-02-21
• [Presentation] Detection of internalization and oligomerization of membrane receptors using coiled-coil fluorescence labeling method (2009)
  • Author(s): Yoshiaki Yano, Kaoru Omae, and Katsumi Matsuzaki
  • Organizer: 第46回ペプチド討論会
  • Place of Presentation: 北九州国際会議場(福岡県)
  • Year and Date: 2009-11-04
• [Presentation] コイルドコイル蛍光ラベル法を用いた生細胞での受容体の可視化解析 (2009)
  • Author(s): 矢野義明
  • Organizer: 大阪大学蛋白質研究所セミナー「膜蛋白質の機能発現メカニズムの解明に向けて-細胞膜上での事象の解析-」
  • Place of Presentation: 大阪 (蛋白質研究所)
  • Year and Date: 2009-09-24
• [Presentation] 受容体内在化を検出する簡便なリガンドスクリーニング法の開発 (2009)
  • Author(s): 矢野義明, 松崎勝巳
  • Organizer: 日本ケミカルバイオロジー学会第4回年会
  • Place of Presentation: 神戸
  • Year and Date: 2009-05-18
• [Book] ナノバイオ技術と最新創薬応用研究「新規蛍光標識法を用いた創薬ターゲット膜受容体のイメージング解析」 (2012)
  • Author(s): 松崎勝巳、矢野義明、他58名
  • Publisher: メディカルドゥ
• [Book] ナノバイオ技術と最新創薬応用研究「新規蛍光標識法を用いた創薬ターゲット膜受容体のイメージング解析」 (2012)
  • Author(s): 松崎勝巳、矢野義明, ほか
  • Total Pages: 288
  • Publisher: メディカルドゥ
• [Book] 薬学分析化学の最前線 (2009)
  • Author(s): 松崎勝巳、矢野義明、他120名
  • Publisher: じほう
• [Remarks]
  • URL: http://www.pharm.kyoto-u.ac.jp/yakkai/
• [Remarks]
  • URL: http://www.pharm.kyoto-u.ac.jp/yakkai/
• [Remarks]
  • URL: http://www.pharm.kyoto-u.ac.jp/yakkai/
• [Remarks] 研究室ホームページ
  • URL: http://www.pharm.kyoto-u.ac.jp/yakkai/