Development of an Active/Inactive Dual Probe Photoaffinity Labeling System for Identification of Natural Product Binding Proteins

• Project/Area Number: 25560400
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biomolecular chemistry
• Research Institution: Tokyo University of Agriculture and Technology
• Principal Investigator: SAKURAI Kaori 東京農工大学, 工学(系)研究科(研究院), 准教授 (50447512)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: フォトアフィニティープローブ / フォトアフィニティーラベリング / 天然物リガンド / OSW-1 / 結合タンパク質

Outline of Final Research Achievements

Identification of binding proteins is critical for understanding the mechanism of action for natural products. In this study, we synthesized an active photoaffinity probe based on an antitumor saponin OSW-1 and corresponding inactive photoaffinity probes to apply the previously developed dual-probe photoaffinity labeling method for the identification of OSW-1 binding proteins. We showed that the active and inactive photoaffinity probes we developed were useful in detecting candidate proteins selective toward the active probe.

Research Products

• [Journal Article] Synthesis of OSW-1 Derivatives by Site-Selective Acylation and Their Biological Evaluation (2014)
  • Author(s): Sakurai, K., Takeshita, T., Hiraizumi, M., Yamada, R.
  • Journal Title: Organic Letters Volume: 16 Issue: 24 Pages: 6318-6321
  • DOI: 10.1021/ol503044j
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Fluorescent analog of OSW-1 and its cellular localization (2014)
  • Author(s): Yamada, R., Takeshita, T., Hiraizumi, M., Shinohe, D., Ohta, Y., Sakurai, K.
  • Journal Title: Bioorganic and Medicinal Chemistry Letters Volume: 24 Issue: 7 Pages: 1839-1842
  • DOI: 10.1016/j.bmcl.2014.02.009
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Comparison of the Reactivity of Carbohydrate Photoaffinity Probes with Different Photoreactive Groups (2014)
  • Author(s): Sakurai, K., Ozawa, S., Yamada, R., Yasui, T., Mizuno, S.
  • Journal Title: ChemBioChem Volume: 15 Issue: 10 Pages: 1399-1403
  • DOI: 10.1002/cbic.201402051
  • Peer Reviewed
• [Presentation] 位置選択的モノアシル化反応を用いたOSW-1不活性体の分子プローブの合成 (2015)
  • Author(s): 平泉将登、武下智哉、山田里佳、櫻井香里
  • Organizer: 第69回有機合成化学協会関東支部シンポジウム
  • Place of Presentation: 横浜国立大学、横浜
  • Year and Date: 2015-05-16
• [Presentation] 位置選択的モノアシル化反応を用いたOSW-1のビオチンプローブの合成と生物活性評価 (2014)
  • Author(s): 武下智哉、平泉将登、 山田里佳、 櫻井香里
  • Organizer: 第4回CSJ化学フェスタ2014
  • Place of Presentation: タワーホール船堀, 東京
  • Year and Date: 2014-10-15
• [Presentation] 位置選択的モノアシル化反応を用いたOSW-1のビオチンプローブの合成と生物活性評価 (2014)
  • Author(s): 武下智哉、平泉将登、 山田里佳、 櫻井香里
  • Organizer: 第67回有機合成化学協会関東支部シンポジウム
  • Place of Presentation: 慶応大学、横浜
  • Year and Date: 2014-05-17
• [Presentation] Fluorescently Labeled OSW-1 and Its Biological Evaluation
  • Author(s): Rika Yamada, Kaori Sakurai
  • Organizer: 第8回ケミカルバイオロジー学会年会
  • Place of Presentation: 東京医科歯科大学（東京）
• [Presentation] Fluorescently Labeled OSW-1 and Its Biological Evaluation
  • Author(s): Rika Yamada, Kaori Sakurai
  • Organizer: 第48回天然物化学談話会
  • Place of Presentation: アヤハレークサイドホテル(大津）
• [Presentation] フォトアフィニティーラベリングを用いた低分子―タンパク質相互作用解析
  • Author(s): Kaori Sakurai
  • Organizer: 第１３回有機合成化学協会関東支部談話会
  • Place of Presentation: 和光純薬工業(株)湯河原研修所（熱海）
  • Invited
• [Presentation] Dual-probe photoaffinity labeling strategy for selective detection of small-molecule binding proteins
  • Author(s): Kaori Sakurai
  • Organizer: 10th Japanese-German Frontiers of Science Symposium
  • Place of Presentation: 京都ブライトンホテル（京都）
  • Invited