Structural analysis of bafilomycin bound to V-ATPase for elucidating its inhibition mechanism based on synthetic chemistry

• Project/Area Number: 15K01821
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Chemical biology
• Research Institution: Osaka University
• Principal Investigator: Tsuchikawa Hiroshi 大阪大学, 理学研究科, 助教 (30460992)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
• Keywords: バフィロマイシン / ATPアーゼ / フッ素標識 / 化学合成 / 構造解析 / 固体NMR / V-ATPase / Bafilomycin / F-NMR

Outline of Final Research Achievements

The structural analysis of bafilomycin (Baf), a specific inhibitor towards vacuolar-type ATPase (V-ATPase), was performed based on synthetic chemistry. First, the preparation of bioactive 19F-labelled Baf required for solid-state NMR measurement was achieved by synthesizing several Baf derivatives and evaluating their V-ATPase inhibitory effect. Next, behavior analysis of active and inactive F-Baf derivatives in model membrane resulted in finding the remarkable difference of their mobility, which may probably relate to the difference of their activity. Moreover, a large-scale preparation of V-ATPase as proteoliposomes was successfully accomplished and the structural analysis of F-Baf bound to V-ATPase was examined by solid-state NMR.

Research Products

• [Journal Article] Stereoselective Construction of Cisoidal Bisspiroacetal Frameworks through Magnesium Coordination of the Bilateral Acetal Oxygen Atoms (2018)
  • Author(s): Yoshifumi Yasukawa, Hiroshi Tsuchikawa, Yasuto Todokoro, and Michio Murata
  • Journal Title: Asian Journal of Organic Chemistry Volume: - Issue: 6 Pages: 1101-1106
  • DOI: 10.1002/ajoc.201800074
  • Peer Reviewed
• [Journal Article] Bafilomycin analogue site-specifically fluorinated at the pharmacophore macrolactone ring has potent vacuolar-type ATPase inhibitory activity (2016)
  • Author(s): Hiroshi Tsuchikawa, Tatsuru Hayashi, Hajime Shibata, Michio Murata, Yoko Nagumo, Takeo Usui
  • Journal Title: Tetrahedron Lett Volume: 57 Issue: 22 Pages: 2426-2429
  • DOI: 10.1016/j.tetlet.2016.04.075
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Elucidation of Bafilomycin-Vacuolar-type ATPase interaction based on solid-state NMR (2018)
  • Author(s): Tatsuru Hayashi, Hiroshi Tsuchikawa, Yuichi Umegawa, Michio Murata, Yoko Nagumo, Takeo Usui
  • Organizer: 日本化学会 第98春季年会
• [Presentation] Investigation of the Interaction between Vacuolar-type ATPase and Bafilomycin by Solid-state NMR (2017)
  • Author(s): Hayashi, T, Tsuchikawa, H, Shibata, H, Murata, M, Nagumo, Y, Usui, T
  • Organizer: 18th Tetrahedron Symposium Asia Edition
  • Int'l Joint Research
• [Presentation] 固体NMRによる配座解析を目指したフッ素標識化バフィロマイシンの合成と生物活性評価 (2016)
  • Author(s): ○林 達・柴田 一・土川 博史・村田 道雄・臼井 健郎
  • Organizer: 日本化学会 第96春季年会
  • Place of Presentation: 同志社大学 京田辺キャンパス
  • Year and Date: 2016-03-24
• [Presentation] Design, synthesis and biological evaluation of fluorinated bafilomycin analogues as an NMR probe to analyze the interaction with vacuolar-type ATPase (2015)
  • Author(s): H. Tsuchikawa, H. Shibata, T. Hayashi, S, Hanashima, N. Matsumori, M. Murata, T. Usui
  • Organizer: PACIFICHEM 2015
  • Place of Presentation: Honolulu, Hawaii, USA
  • Year and Date: 2015-12-15
  • Int'l Joint Research