Controlling the Rigidity of Kinesin-Propelled Microtubules in an In Vitro Gliding Assay Using the Deep-Sea Osmolyte Trimethylamine N-Oxide

Arif Md Rashedul Kabir; Tasrina Munmun; Tomohiko Hayashi; Satoshi Yasuda; Atsushi P Kimura; Masahiro Kinoshita; Takeshi Murata; Kazuki Sada; Akira Kakugo

doi:10.1021/acsomega.1c06699

Controlling the Rigidity of Kinesin-Propelled Microtubules in an In Vitro Gliding Assay Using the Deep-Sea Osmolyte Trimethylamine N-Oxide

ACS Omega. 2022 Jan 24;7(4):3796-3803. doi: 10.1021/acsomega.1c06699. eCollection 2022 Feb 1.

Authors

Arif Md Rashedul Kabir¹, Tasrina Munmun², Tomohiko Hayashi³, Satoshi Yasuda^{4

5}, Atsushi P Kimura^{1

6}, Masahiro Kinoshita^{3

4

5}, Takeshi Murata^{4

5}, Kazuki Sada^{1

2}, Akira Kakugo^{1

2}

Affiliations

¹ Faculty of Science, Hokkaido University, Sapporo 060-0810, Japan.
² Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo 060-0810, Japan.
³ Institute of Advanced Energy, Kyoto University, Uji, Kyoto 611-0011, Japan.
⁴ Graduate School of Science, Chiba University, 1-33 Yayoi-cho, Inage, Chiba 263-8522, Japan.
⁵ Membrane Protein Research and Molecular Chirality Research Centers, Chiba University, 1-33 Yayoi-cho, Inage, Chiba 263-8522, Japan.
⁶ Graduate School of Life Science, Hokkaido University, Sapporo 060-0810, Japan.

Abstract

The biomolecular motor protein kinesin and its associated filamentous protein microtubule have been finding important nanotechnological applications in the recent years. Rigidity of the microtubules, which are propelled by kinesin motors in an in vitro gliding assay, is an important metric that determines the success of utilization of microtubules and kinesins in various applications, such as transportation, sensing, sorting, molecular robotics, etc. Therefore, regulating the rigidity of kinesin-propelled microtubules has been critical. In this work, we report a simple strategy to regulate the rigidity of kinesin-propelled microtubules in an in vitro gliding assay. We demonstrate that rigidity of the microtubules, propelled by kinesins in an in vitro gliding assay, can be modulated simply by using the natural osmolyte trimethylamine N-oxide (TMAO). By varying the concentration of TMAO in the gliding assay, the rigidity of microtubules can be modulated over a wide range. Based on this strategy, we are able to reduce the persistence length of microtubules, a measure of microtubule rigidity, ∼8 fold by using TMAO at the concentration of 1.5 M. Furthermore, we found that the decreased rigidity of the kinesin-propelled microtubules can be restored upon elimination of TMAO from the in vitro gliding assay. Alteration in the rigidity of microtubules is accounted for by the non-uniformity of the force applied by kinesins along the microtubules in the presence of TMAO. This work offers a facile strategy to reversibly regulate the rigidity of kinesin-propelled microtubules in situ, which would widen the applications of the biomolecular motor kinesin and its associated protein microtubule in various fields.