Development of labeling/purifying/detecting methods using a reactive affinity tag

• Project/Area Number: 26560439
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biomolecular chemistry
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: DODO KOSUKE 国立研究開発法人理化学研究所, 袖岡有機合成化学研究室, 専任研究員 (20415243)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
• Keywords: 有機化学 / タンパク質 / ペプチド / アフィニティー精製 / アフィニティーラベリング / 蛍光標識 / 金属錯体

Outline of Final Research Achievements

To identify the target proteins and binding sites of bioactive compounds, we planned to develop “reactive affinity tag”, which can label/purify/detect binding proteins of bioactive compounds. As a result, we successfully installed another function into β-ketoamide or O-nitrobenzoxadiazole (NBD) unit. Finally, we developed a reactive affinity tag, which enabled us to label/detect the binding proteins of bioactive compounds and purify/identify the labeled peptides.

Research Products

• [Journal Article] Unique features of chiral palladium enolates derived from β-ketoamide: Structure and catalytic asymmetric Michael and fluorination reactions (2015)
  • Author(s): Kenji Hayamizu, Naoki Terayama, Daisuke Hashizume, Kosuke Dodo, and Mikiko Sodeoka
  • Journal Title: Tetrahedron Volume: 71 Issue: 37 Pages: 6594-6601
  • DOI: 10.1016/j.tet.2015.07.002
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Pd enolate錯体を用いたアフィニティー精製法の開発 (2017)
  • Author(s): 早水健二、どど孝介、江上寛通、淺沼三和子、袖岡幹子
  • Organizer: 日本薬学会第１３７年会
  • Place of Presentation: 仙台国際センター（宮城県・仙台）
  • Year and Date: 2017-03-24
• [Presentation] β-ケトアミドのPdエノラートを用いた触媒的不斉付加反応の開発 (2017)
  • Author(s): 寺山直樹、早水健二、橋爪大輔、どど孝介、袖岡 幹子
  • Organizer: 日本薬学会第１３７年会
  • Place of Presentation: 仙台国際センター（宮城県・仙台）
  • Year and Date: 2017-03-24
• [Presentation] Turn-ON蛍光アフィニティー標識法を用いた生物活性化合物の標的タンパク質解析 (2017)
  • Author(s): 淺沼三和子、どど孝介、大金賢司、袖岡 幹子
  • Organizer: 日本薬学会第１３７年会
  • Place of Presentation: 仙台国際センター（宮城県・仙台）
  • Year and Date: 2017-03-24
• [Presentation] Turn-ON fluorescent affinity labeling using small bifunctional O-NBD unit (2016)
  • Author(s): Kosuke Dodo
  • Organizer: 2016 Queenstown Molecular Biology Meetings
  • Place of Presentation: Shanghai, China
  • Year and Date: 2016-03-17
  • Int'l Joint Research / Invited
• [Presentation] 蛍光turn-ON型ラベル化法と質量分析法を用いた生物活性化合物の標的タンパク質解析 (2015)
  • Author(s): 淺沼三和子、どど孝介、藤本直子、袖岡幹子
  • Organizer: BMB2015（第38回日本分子生物学会年会・第88回日本生化学会大会 合同大会）
  • Place of Presentation: 神戸ポートアイランド（兵庫・神戸市）
  • Year and Date: 2015-12-01
• [Presentation] Unique features of chiral palladium enolates derived from β-ketoamide: Structure and catalytic asymmetric Michael and fluorination reactions (2015)
  • Author(s): Kenji Hayamizu, Naoki Terayama, Daisuke Hashizume, Kosuke Dodo, and Mikiko Sodeoka
  • Organizer: 16th Tetrahedron Symposium Asia edition
  • Place of Presentation: Shanghai, China
  • Year and Date: 2015-11-10
  • Int'l Joint Research
• [Presentation] Turn-ON fluorescent affinity labeling using small bifunctional O-NBD unit (2015)
  • Author(s): Kosuke Dodo, Takao Yamaguchi, Miwako Asanuma, Shuichi Nakanishi, Yohei Saito, Masateru Okazaki, Mikiko Sodeoka
  • Organizer: European Chemical Biology Society (ECBS) & International Chemical Biology Society (ICBS) Joint Meeting
  • Place of Presentation: Berlin, Germany
  • Year and Date: 2015-10-07
  • Int'l Joint Research
• [Presentation] ネクローシス制御機構解明を目指したケミカルバイオロジー研究：蛍光turn-ON型アフィニティーラベル化法の開発 (2015)
  • Author(s): どど孝介
  • Organizer: 第15回日本蛋白質科学会年会
  • Place of Presentation: あわぎんホール（徳島・徳島市）
  • Year and Date: 2015-06-24
  • Invited
• [Presentation] 蛍光turn-ON型ラベル化法を用いた生物活性化合物の標的タンパク質の解析 (2014)
  • Author(s): 淺沼三和子、どど孝介、山口卓男、中西修一、斎藤洋平、岡﨑正晃、袖岡幹子
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 国立京都国際会館（京都）
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] ニトロベンゾオキサジアゾールを用いる蛍光turn-ON型アフィニティーラベル化法の開発 (2014)
  • Author(s): 山口卓男、淺沼三和子、中西修一、斎藤洋平、岡﨑正晃、どど孝介、袖岡幹子
  • Organizer: 第12回 次世代を担う有機化学シンポジウム
  • Place of Presentation: 日本薬学会長井記念ホール（東京）
  • Year and Date: 2014-05-23 – 2014-05-24