High-Resolution Imaging of a Single Gliding Protofilament of Tubulins by HS-AFM

Jakia Jannat Keya, Daisuke Inoue, Yuki Suzuki, Toshiya Kozai, Daiki Ishikuro, Noriyuki Kodera, Takayuki Uchihashi, Arif Md Rashedul Kabir, Masayuki Endo, Kazuki Sada, Akira Kakugo

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Abstract

In vitro gliding assay of microtubules (MTs) on kinesins has provided us with valuable biophysical and chemo-mechanical insights of this biomolecular motor system. Visualization of MTs in an in vitro gliding assay has been mainly dependent on optical microscopes, limited resolution of which often render them insufficient sources of desired information. In this work, using high speed atomic force microscopy (HS-AFM), which allows imaging with higher resolution, we monitored MTs and protofilaments (PFs) of tubulins while gliding on kinesins. Moreover, under the HS-AFM, we also observed splitting of gliding MTs into single PFs at their leading ends. The split single PFs interacted with kinesins and exhibited translational motion, but with a slower velocity than the MTs. Our investigation at the molecular level, using the HS-AFM, would provide new insights to the mechanics of MTs in dynamic systems and their interaction with motor proteins.

Original languageEnglish
Article number6166
JournalScientific reports
Volume7
Issue number1
DOIs
Publication statusPublished - Dec 1 2017

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Keya, J. J., Inoue, D., Suzuki, Y., Kozai, T., Ishikuro, D., Kodera, N., ... Kakugo, A. (2017). High-Resolution Imaging of a Single Gliding Protofilament of Tubulins by HS-AFM. Scientific reports, 7(1), [6166]. https://doi.org/10.1038/s41598-017-06249-1