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We thank Ken'ya Furuta (University of Tokyo) for kind help in MT gliding assay and generous gift of Mark2, and Roger Y. Tsien (Howard Hughes Medical Institute, UCSD) for providing a cDNA for mCherry.
Right top: schematic of flow-cell-based kinesin gliding assay.
The gliding assay chambers were prepared by using hydrophilic coverslips.
The de novo KIF1A mutants were nonmotile in the microtubule gliding assay.
(A ) Microtubule gliding assay on a Drosophila kinesin-1-coated surface.
The microtubule gliding assay was designed as described before (Nitzsche et al., 2010).
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Thus, gliding motility works similarly to microtubule gliding assays, in which surface-immobilized dyneins glide microtubules with their plus-end tips in the lead.
A coarse-grained computational model of gliding assays was developed and found to recapitulate the experiments.
To understand how diverse kinesins attached to the same cargo coordinate their movement, we carried out microtubule gliding assays using pairwise mixtures of motors from the kinesin-1, -2, -3, -5, and -7 families engineered to have identical run lengths and surface attachments.
Several observations made with in vitro motility assays have shown a torque component to the working stroke, dubbed "twirling", inferred from rotations of the actin filament in gliding assays [84], [85].
Microtubule gliding assays and microtubule binding and release assays were performed as described (Huang et al., 2012).
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