Abstract
AbstractIn this study, we report differences in the observed gliding speed of microtubules dependent on the choice of bovine casein used as a surface passivator. We observed differences in both speed and support of microtubules in each of the assays. Whole casein, comprised of [alpha]~s1~, [alpha]~s2~, [beta], and [kappa] casein, supported motility and averaged speeds of 966 ± 7 nm/s. Alpha casein can be purchased as a combination of s1 and s2 and supported gliding motility and average speeds of 949 ± 4 nm/s. Beta casein did not support motility very well and averaged speeds of 870 ± 30 nm/s. Kappa casein supported motility very poorly and we were unable to obtain an average speed. Finally, we observed that mixing alpha, beta, and kappa casein with the proportions found in bovine whole casein supported motility and averaged speeds of 966 ± 7 nm/s.
Highlights
Kinesin-1 is an ATPase that converts chemical energy to mechanical work
The other method is a gliding motility assay where kinesins are fixed to a glass slide and microtubules flow on top of a layer of kinesin [9,10,11]
In the gliding motility assay, motility is sustained by first passivating the glass to prevent kinesin’s motor domains from becoming inactive when interacting with untreated glass
Summary
Kinesin-1 (hereafter referred to as kinesin) is an ATPase that converts chemical energy to mechanical work. It travels along microtubules in one direction and can carry with it various cellular items [1,2,3,4,5]. Microtubules are fixed to a coverglass and individual kinesin motion are observed either by single-fluorophore tracking, or by attaching beads to kinesin [6,7,8]. The other method is a gliding motility assay where kinesins are fixed to a glass slide and microtubules flow on top of a layer of kinesin [9,10,11]. In the gliding motility assay, motility is sustained by first passivating the glass to prevent kinesin’s motor domains from becoming inactive when interacting with untreated glass
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