Structural and physical properties of the clutch molecular complex in motile cells.

• Project/Area Number: 26251006
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Nara Institute of Science and Technology
• Principal Investigator: Hakoshima Toshio 奈良先端科学技術大学院大学, バイオサイエンス研究科, 教授 (00164773)
• Co-Investigator(Kenkyū-buntansha): 稲垣 直之 奈良先端科学技術大学院大学, バイオサイエンス研究科, 教授 (20223216)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥40,040,000 (Direct Cost: ¥30,800,000、Indirect Cost: ¥9,240,000); Fiscal Year 2016: ¥11,700,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥2,700,000); Fiscal Year 2015: ¥19,630,000 (Direct Cost: ¥15,100,000、Indirect Cost: ¥4,530,000); Fiscal Year 2014: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000)
• Keywords: 構造生物学 / 生化学 / 生物物理学 / 分子細胞生物学 / タンパク質 / 相互作用 / 構造変化 / 制御機構 / 細胞生物学 / 分子生物学 / 構造 / X線 / 神経 / 軸索

Outline of Final Research Achievements

The traction forces underlying cellular motility is regulated in part by the phosphorylation-induced modulation of coupling efficiency of the "clutch" molecule Shootin1 between F-actin and adhesion molecule L1-CAM, which bound the extracellular matrix. To establish the molecular basis of the regulatory mechanism, we analyzed by physical methods the structural and physical properties of Shootin1 and its physical interactions with L1-CAM and other regulatory proteins using purified protein samples. We found that three α-helical regions at the N-terminal half of Shootin1 and the phosphorylation sites existing at the first and second helical regions. The helical regions mediate Shootin1dimerization in solution and the phosphorylation at the N-terminal helical region induces formation of a tetramer or even higher oligomers to directly bind the cytoplasmic region of L1-CAM.

Research Products

• [Journal Article] Grip and slip of L1-CAM on adhesive substrates direct growth cone haptotaxis. Proc. Natl. (2018)
  • Author(s): Abe, K., Katsuno, H., Toriyama, M., Baba, K., Mori, T., Hakoshima, T., Kanemura, Y., Watanabe, R. and Inagaki, N.
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 115 Issue: 11 Pages: 2764-2769
  • DOI: 10.1073/pnas.1711667115
  • Peer Reviewed / Open Access
• [Journal Article] Real-time TIRF observation of vinculin recruitment to stretched α-catenin by AFM. (2018)
  • Author(s): Maki, K., Han, S. -U., Hirano, Y., Yonemura, S. Hakoshima, T. and Adachi, T.
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 1575-1575
  • DOI: 10.1038/s41598-018-20115-8
  • NAID: 120006517895
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Structural basis of thalidomide enantiomer binding to cereblon. (2018)
  • Author(s): Mori, T., Itoh, T., Liu, S., Ando, H., Sakamoto, S., Yamaguchi, Y., Tokunaga, E., Shibata, N., Handa, H. and Hakoshima, T.
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 1294-1294
  • DOI: 10.1038/s41598-018-19202-7
  • Peer Reviewed / Open Access
• [Journal Article] The force‐sensing device region of α‐catenin is an intrinsically disordered segment in the absence of intramolecular stabilization of the autoinhibitory form (2018)
  • Author(s): Hirano, Y., Amano, Y., Yonemura, S. and Hakoshima, T.
  • Journal Title: Genes to Cells Volume: 23 Issue: 5 Pages: 16-16
  • DOI: 10.1111/gtc.12578
  • Peer Reviewed / Open Access
• [Journal Article] Mechano-adaptive sensory mechanism of α-catenin under tension (2016)
  • Author(s): Koichiro Maki, Sung-Woong Han, Yoshinori Hirano, Shigenobu Yonemura, Toshio Hakoshima & Taiji Adachi
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 24878-24878
  • DOI: 10.1038/srep24878
  • NAID: 120006337976
  • Peer Reviewed / Open Access
• [Journal Article] Structural basis for autoinhibition and its relief of MOB1 in the Hippo pathway (2016)
  • Author(s): Sun-Yong Kim, Yuka Tachioka, Tomoyuki Mori & Toshio Hakoshima
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 28488-28488
  • DOI: 10.1038/srep28488
  • NAID: 120005841618
  • Peer Reviewed / Open Access
• [Journal Article] Structure of the free form of the N-terminal VH1 domain of monomeric α-catenin. (2015)
  • Author(s): Shibahara T, Hirano Y, Hakoshima T.
  • Journal Title: FEBS Letter Volume: 589 Issue: 15 Pages: 1754-1760
  • DOI: 10.1016/j.febslet.2015.05.053
  • Peer Reviewed / Open Access
• [Journal Article] MT1-MMP recognition by ERM proteins and its implication in CD44 shedding. (2015)
  • Author(s): Shin-ichi Terawaki, Ken Kitano, Miki Aoyama, Tomoyuki Mori, Toshio Hakoshima
  • Journal Title: Genes to Cells Volume: 20(10) Issue: 10 Pages: 847-859
  • DOI: 10.1111/gtc.12276
  • Peer Reviewed / Open Access
• [Book] Protein modification for crystallization. in Advanced Methods in Structural Biology (eds T. Senda, K. Maenaka) (2016)
  • Author(s): Hakoshima, T.
  • Total Pages: 340
  • Publisher: Springer