Kinesin-1 Expressed in Insect Cells Improves Microtubule in Vitro Gliding Performance, Long-Term Stability and Guiding Efficiency in Nanostructures

IEEE Transactions on Nanobioscience
Till KortenStefan Diez

Abstract

The cytoskeletal motor protein kinesin-1 has been successfully used for many nanotechnological applications. Most commonly, these applications use a gliding assay geometry where substrate-attached motor proteins propel microtubules along the surface. So far, this assay has only been shown to run undisturbed for up to 8 h. Longer run times cause problems like microtubule shrinkage, microtubules getting stuck and slowing down. This is particularly problematic in nanofabricated structures where the total number of microtubules is limited and detachment at the structure walls causes additional microtubule loss. We found that many of the observed problems are caused by the bacterial expression system, which has so far been used for nanotechnological applications of kinesin-1. We strive to enable the use of this motor system for more challenging nanotechnological applications where long-term stability and/or reliable guiding in nanostructures is required. Therefore, we established the expression and purification of kinesin-1 in insect cells which results in improved purity and--more importantly--long-term stability > 24 h and guiding efficiencies of > 90% in lithographically defined nanostructures.

Citations

Mar 23, 2017·Chemical Society Reviews·H Hess, Jennifer L Ross
Aug 11, 2018·Proceedings of the National Academy of Sciences of the United States of America·Aniruddha MitraStefan Diez
Mar 7, 2021·International Journal of Molecular Sciences·Anne SeifertAndreas Hermann
Sep 13, 2019·Nano Letters·Mohammad A RahmanAlf Månsson
Oct 31, 2018·Accounts of Chemical Research·Henry Hess, Gadiel Saper
Sep 12, 2019·Chemical Reviews·Gadiel Saper, Henry Hess
Jun 20, 2020·Langmuir : the ACS Journal of Surfaces and Colloids·Neda M Bassir KazeruniHenry Hess
Jan 21, 2022·Nano Letters·Cordula ReutherStefan Diez

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