title: Kinesin-1 Expressed in Insect Cells Improves Microtubule in Vitro Gliding Performance, Long-Term Stability and Guiding Efficiency in Nanostructures.
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.
authors.forename: Till
authors.initials: T
authors.lastname: Korten
authors.forename: Samata
authors.initials: S
authors.lastname: Chaudhuri
authors.forename: Elena
authors.initials: E
authors.lastname: Tavkin
authors.forename: Marcus
authors.initials: M
authors.lastname: Braun
authors.forename: Stefan
authors.initials: S
authors.lastname: Diez
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D029721:Drosophila Proteins / Q000494:pharmacology*; D016547:Kinesins / Q000494:pharmacology*; D008870:Microtubules / Q000187:drug effects* / Q000502:physiology*; D008954:Models, Biological; D049329:Nanostructures / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods*; D013552:Swine; D014404:Tubulin / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; D014404:Tubulin; C515209:Khc protein, Drosophila; D016547:Kinesins