Development of photoaffinity selection system for identification of low abundance proteins

• Project/Area Number: 15K05559
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Bio-related chemistry
• Research Institution: Tokyo University of Agriculture and Technology
• Principal Investigator: Kaori Sakurai 東京農工大学, 工学(系)研究科(研究院), 准教授 (50447512)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2017: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2015: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: フォトアフィニティープローブ / 生物活性分子 / 結合タンパク質 / 金ナノ粒子 / 結合タンパク質探索法 / 標的タンパク質 / 光反応基 / フォトアフィニティーラベリング / フォトアフィニティラベリング / アフィニティー精製

Outline of Final Research Achievements

Development of efficient target identification methods is important for accelerating the discovery of new biological pathways and drug targets. We have designed and synthesized gold-nanoparticle based affinity labeling probes, which display bioactive ligands and photoreactive groups for catch and release of binding proteins in one-pot. Gold nanoparticles are suitable as probe scaffolds due to its capability to covalently immobilize organic compounds in a multivalent fashion, the water dispersibility and the high density, which enable facie centrifugal separation. We demonstrated that our gold-nanoparticle probes can crosslink the binding proteins of high μM affinity with high selectivity at nM probe concentration.

Research Products

• [Int'l Joint Research] カロリンスカ研究所(スウェーデン)
• [Journal Article] Synthesis of alkyne-tagged and biotin-tagged Sortin1 as novel photoaffinity probes (2018)
  • Author(s): Komatsu, R. Yamaguchi, T. Kobayashi, N. Ozeki, Y. Sakurai, K.
  • Journal Title: Bioorganic & Medicinal Chemistry Letters Volume: 28 Issue: 9 Pages: 1562-1565
  • DOI: 10.1016/j.bmcl.2018.03.060
  • Peer Reviewed
• [Journal Article] Synthesis of a fluorescent photoaffinity probe of OSW-1 by site-selective acylation of an inactive congener and biological evaluation (2017)
  • Author(s): Sakurai, K., Hiraizumi, M., Isogai, N., Komatsu, R., Shibata, T., Ohta, Y.
  • Journal Title: Chemical Communications Volume: 53 Issue: 20 Pages: 517-520
  • DOI: 10.1016/j.bmcl.2016.08.053
  • Peer Reviewed
• [Journal Article] Gold Nanoparticle-Based Multivalent Carbohydrate Probes: Selective Photoaffinity Labeling of Carbohydrate-Binding Proteins (2016)
  • Author(s): Sakurai, K., Hatai, Y., Okada, A.
  • Journal Title: Chem. Sci. Volume: 7 Issue: 1 Pages: 702-706
  • DOI: 10.1039/c5sc03275j
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] 生物活性分子の作用機構解明に向けた標的タンパク質探索法 (2018)
  • Author(s): 櫻井香里
  • Organizer: Asian Chemical Probe Research Hub Symposium天然物有機化学・天然物合成とケミカルバイオロジー
  • Invited
• [Presentation] 生物活性分子の作用機構解明に向けた標的タンパク質探索法 (2018)
  • Author(s): 櫻井香里
  • Organizer: 日本化学会 第98春季年会 女性科学者が拓く生命化学 特別企画公演
  • Invited
• [Presentation] 生物活性分子の作用機序解明に向けた標的タンパク質探索法の開発 (2017)
  • Author(s): 櫻井香里
  • Organizer: 新規素材探索研究会第16回セミナー
  • Invited
• [Presentation] Development of gold nanoparticle-based photoaffinity probes displaying bioactive small-molecule ligands (2017)
  • Author(s): Amane Kato, Keisuke Adachi, Yuki Hatai, Kaori Sakurai,
  • Organizer: 第９７回日本化学会春季年会
  • Place of Presentation: 慶應大学日吉キャンパス（神奈川県横浜市）
• [Presentation] Development of gold nanoparticle-based multivalent carbohydrate probes for selective photoaffinity labeling and enrichment of carbohydrate-binding proteins (2015)
  • Author(s): Yuki Hatai
  • Organizer: 第34回日本糖質学会年会
  • Place of Presentation: 東京大学安田講堂（東京都文京区）
  • Year and Date: 2015-08-02
• [Remarks]
  • URL: http://web.tuat.ac.jp/~sakurai/