High-speed AFM study on functional modulation of kinesin caused by structural defects of mictoruble

2020 Fiscal Year Final Research Report

• Project/Area Number: 18H01837
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 28040:Nanobioscience-related
• Research Institution: Nagoya University
• Principal Investigator: Uchihashi Takayuki 名古屋大学, 理学研究科, 教授 (30326300)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: 一分子計測 / 微小管 / キネシン / タンパク質 / 原子間力顕微鏡

Outline of Final Research Achievements

In this study, we have developed an in-line force-curve mode, which is an advanced molecular manipulation function of high-speed atomic force microscopy (AFM), allowing us to form single tubulin dimer defects in microtubules with high controllability and to obtain force-distance curves in realtime. This technique enabled us to observe the self-healing process of structural defects created on a microtubule and the translational motility of kinesin molecules around the defects, and to quantify the binding energy between tubulins. Furthermore, we have extended this technique to the fast force mapping mode. Also, by observing the kinesin sliding motion in bent microtubules, we found that the kinesin translational velocity is significantly reduced in the bent region.

Free Research Field

生物物理学

Academic Significance and Societal Importance of the Research Achievements

本研究で開発された手法は、微小管の機械特性によるMAPsの機能変調を解析できる唯一手法であり、今後微小管に限らず様々なタンパク質の一分子構造操作や機能解析が可能になることから、生物学の重要な計測ツールとなり学術的意義は高い。