Crosslinking and miniaturization of protein labeling method in living cells based on coiled-coil formation

• Project/Area Number: 20790033
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Physical pharmacy
• Research Institution: Kyoto University
• Principal Investigator: YANO Yoshiaki Kyoto University, 薬学研究科, 助教 (60402799)
• Project Period (FY): 2008 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2009: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2008: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: 蛍光タンパク質 / コイルドコイル / イメージング / Gタンパク質共役型受容体 / クロスリンク / ATTO488 / ATTO700 / factorXa / 受容体 / プロテアーゼ / ローダミン / フルオレセイン

Research Abstract

Live-cell imaging of target proteins by using genetic fusion with fluorescent proteins is widely used in basic life-science research. We performed crosslinking and further miniaturization of a recently developed labeling technology (coiled-coil labeling method), which has a smaller size compared with fluorescent protein.

Research Products

• [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 labeling methods for live-cell imaging of membrane proteins. (2009)
  • Author(s): Yano Y, Matsuzaki K
  • Journal Title: Biochim. Biopys. Acta 1788 Pages: 2124-2131
  • NAID: 120001645079
  • 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] Coiled-coil tag-probe system for quick labeling of membrane receptors in living cell. (2008)
  • Author(s): Yano Y, Yano A, Oishi S, Sugimoto Y, Tsujimoto G, Fujii N, Matsuzaki K
  • Journal Title: ACS Chemical Biology 3 Pages: 341-345
  • Peer Reviewed
• [Journal Article] Coiled-coil tag-probe system for quick labeling of membrane receptors in livina cell (2008)
  • Author(s): Yano Y, Yano A, Oishi S, Sugimoto Y, Tsujimoto G, Fujii N, Matsuzaki K
  • Journal Title: ACS Chemical Biology 3 Pages: 345-345
  • Peer Reviewed
• [Journal Article] Intracellular third loop-C-terminal tail interaction in prostaglandin EP3beta receptor (2008)
  • Author(s): Yano Y, Shimbo T, Sugimoto Y, Matsuzaki K
  • Journal Title: Biochemical and Biophysical Research Communications 371 Pages: 846-849
  • Peer Reviewed
• [Presentation] 新規小分子蛍光ラベル法による生細胞膜での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(岡山大学)
  • Year and Date: 2010-03-28
• [Presentation] 新規小分子蛍光ラベル法による生細胞膜での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: 岡山大学
  • Year and Date: 2010-03-26
• [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 ANALYSIS 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 Centor)
  • Year and Date: 2010-02-21
• [Presentation] 新規タグープローブ法で見る膜受容体の生細胞での挙動 (2009)
  • Author(s): 矢野義明, 松崎勝巳
  • Organizer: 日本薬学会第129年会
  • Place of Presentation: 京都国際会館
  • Year and Date: 2009-03-26
• [Presentation] Visualizing receptor interanlization using a new fluorescence labeling method (2009)
  • Author(s): 矢野義明、松崎勝巳
  • Organizer: Biophysical society 53rd annual meeting
  • Place of Presentation: Boston USA
  • Year and Date: 2009-03-01
• [Presentation] Coiled-coil tag-probe system for quick labeling of membrane receptors in living cells (2008)
  • Author(s): 矢野義明, 崎勝巳
  • Organizer: International symposium on integrated medicinal science (IMS 2008)
  • Place of Presentation: 京都薬科大学
  • Year and Date: 2008-12-13
• [Presentation] Visualizing internalization of membrane receptors using new fluorescence labeling method (2008)
  • Author(s): 矢野義明、松崎勝巳
  • Organizer: 第45回ペプチド討論会
  • Place of Presentation: 東京タワーホール船堀
  • Year and Date: 2008-10-31
• [Book] 薬学分析化学の最前線 (2009)
  • Author(s): 松崎勝巳, 矢野義明
  • Publisher: じほう
• [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/
• [Remarks]
  • URL: http://www.pharm.kyoto-u.ac.jp/yakkai/