Development of high-speed atomic force microscope with minimum load for observing protein molecules

• Project/Area Number: 17K19042
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Research Field: Nano/Micro science and related fields
• Research Institution: Fukuoka University
• Principal Investigator: YAMAMOTO Daisuke 福岡大学, 理学部, 教授 (80377902)
• Co-Investigator(Kenkyū-buntansha): 武藤 梨沙 福岡大学, 理学部, 助教 (10622417)
• Project Period (FY): 2017-06-30 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000); Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2017: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: 走査プローブ顕微鏡 / ナノバイオ / 蛋白質 / 原子間力顕微鏡 / 生物物理

Outline of Final Research Achievements

A novel method of atomic force microscopy (AFM) was developed to enable the observations of biological molecules with very low loading force. This method was introduced to high-speed AFM and protein molecules were examined. The force acting between the probe tip and the sample surface was estimated. It was found that the loading force is significantly small compared to the conventional high-speed AFM. It was confirmed that this novel method significantly reduces the invasiveness to the protein GroEL compared to the conventional high-speed AFM. Furthermore, surface structure unprecedented by conventional AFM observations was found on membrane protein bacteriorhodopsin.

Academic Significance and Societal Importance of the Research Achievements

タンパク質は柔軟な分子機械であるため、そのありのままの動態を解析するためには、測定がタンパク質に与える擾乱を可能な限り低減させる必要がある。本研究で開発したAFM測定法は、探針－試料間相互作用の大きさを極めて小さくすることができる。そのため、従来のAFM測定法と比較して、タンパク質が本来供えている構造動態を乱すことなく測定することができると考えられる。本手法を拡張することで、将来的に多様なAFM測定法が開発されることにつながることも期待される。

Research Products

• [Journal Article] Electron transport pathways in isolated chromoplasts from Narcissus pseudonarcissus L. (2019)
  • Author(s): Grabsztunowicz Magda、Mulo Paula、Baymann Frauke、Mutoh Risa、Kurisu Genji、Setif Pierre、Beyer Peter、Krieger‐Liszkay Anja
  • Journal Title: The Plant Journal Volume: - Issue: 2 Pages: 245-256
  • DOI: 10.1111/tpj.14319
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] X-ray structure of an asymmetrical trimeric ferredoxin-photosystem I complex (2018)
  • Author(s): Kubota-Kawai Hisako、Mutoh Risa、Shinmura Kanako、Setif Pierre、Nowaczyk Marc M.、Rogner Matthias、Ikegami Takahisa、Tanaka Hideaki、Kurisu Genji
  • Journal Title: Nature Plants Volume: 4 Issue: 4 Pages: 218-224
  • DOI: 10.1038/s41477-018-0130-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] X-ray crystallographic and high-speed AFM studies of peroxiredoxin 1 from Chlamydomonas reinhardtii (2018)
  • Author(s): Charoenwattanasatien Ratana、Tanaka Hideaki、Zinzius Karen、Hochmal Ana K.、Mutoh Risa、Yamamoto Daisuke、Hippler Michael、Kurisu Genji
  • Journal Title: Acta Crystallographica Section F Structural Biology Communications Volume: 74 Issue: 2 Pages: 86-91
  • DOI: 10.1107/s2053230x17018507
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Association of ferredoxin:NADP+ oxidoreductase with the photosynthetic apparatus modulates electron transfer in Chlamydomonas reinhardtii. (2017)
  • Author(s): Mosebach, L., Heilmann, C., Mutoh, R., Gaebelein, P., Steinbeck, J., Happe, T., Ikegami, T., Hanke, G., Kurisu, G., Hippler, M.
  • Journal Title: Photosynth. Res. Volume: 134 Issue: 3 Pages: 291-306
  • DOI: 10.1007/s11120-017-0408-5
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Dynamics and energetics of cyanobacterial photosystem I:ferredoxin complexes in different redox states (2017)
  • Author(s): Setif Pierre、Mutoh Risa、Kurisu Genji
  • Journal Title: Biochimica et Biophysica Acta (BBA) - Bioenergetics Volume: 1858 Issue: 7 Pages: 483-496
  • DOI: 10.1016/j.bbabio.2017.04.001
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 強光下における光合成膜内タンパク質の動的挙動解析 (2019)
  • Author(s): 太田明香音、武藤梨沙、得津隆太郎、皆川純、山本大輔
  • Organizer: 第42回日本分子生物学会年会
• [Presentation] ホウレンソウグラナ膜における光化学系II分子間相互作用の高速AFMによる解析 (2019)
  • Author(s): 山本大輔
  • Organizer: 第57回日本生物物理学会年会
• [Presentation] ホウレンソウ由来ストロマラメラに内在するF0 c-リングの原子間力顕微鏡による観察 (2018)
  • Author(s): 山本大輔、武藤梨沙
  • Organizer: 第56回日本生物物理学会年会
• [Presentation] 光化学系I-フェレドキシン複合体の構造解析 (2018)
  • Author(s): 武藤梨沙
  • Organizer: NMR若手研究会
  • Invited
• [Presentation] X-ray structure of the type-I reaction center from Heliobacterium modesticaldum at 3.2 Å resolution (2018)
  • Author(s): T. Nakaniwa, R. Mutoh, K. Fushimi, A. Yasuda, T. Mizoguchi, H. Tamiaki, C. Azai, H. Tanaka, S. Itoh, H. Oh-oka, G. Kurisu
  • Organizer: 第56回日本生物物理学会年会
  • Invited
• [Presentation] The dynamics of photosystem 2 and light-harvesting complex 2 in spinach grana membrane revealed by high-speed AFM (2017)
  • Author(s): Risa Mutoh, Takahiro Iida, Daisuke Yamamoto
  • Organizer: 第55回日本生物物理学会年会