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Microtubule detachment in gliding motility assays limits the performance of kinesin-driven molecular shuttles.

The creation of complex active nanosystems integrating cytoskeletal filaments propelled by surface-adhered motor proteins often relies on the filaments ability to glide over up to meter-long distances. While theoretical considerations support this ability, we show that microtubule detachment (either spontaneous or triggered by a microtubule crossing event) is a non-negligible phenomenon that has been overlooked until now. The average gliding distance before spontaneous detachment was measured to be 30±10 mm for a functional kinesin-1 density of 500 μm-2 and 9±4 mm for a functional kinesin-1 density of 100 μm-2 at 1 mM ATP. Even microtubules longer than 3 μm detached, suggesting that spontaneous detachment is not caused by the stochastic absence of motors or their stochastic release due to a limited run length.