Kinesin-1 activity recorded in living cells with a precipitating dye

Simona Angerani, Eric Lindberg, Nikolai Klena, Christopher K. E. Bleck, Charlotte Aumeier () and Nicolas Winssinger ()
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Simona Angerani: University of Geneva
Eric Lindberg: University of Geneva
Nikolai Klena: University of Geneva
Christopher K. E. Bleck: National Institutes of Health
Charlotte Aumeier: University of Geneva
Nicolas Winssinger: University of Geneva

Nature Communications, 2021, vol. 12, issue 1, 1-10

Abstract: Abstract Kinesin-1 is a processive motor protein that uses ATP-derived energy to transport a variety of intracellular cargoes toward the cell periphery. The ability to visualize and monitor kinesin transport in live cells is critical to study the myriad of functions associated with cargo trafficking. Herein we report the discovery of a fluorogenic small molecule substrate (QPD-OTf) for kinesin-1 that yields a precipitating dye along its walking path on microtubules (MTs). QPD-OTf enables to monitor native kinesin-1 transport activity in cellulo without external modifications. In vitro assays show that kinesin-1 and MTs are sufficient to yield fluorescent crystals; in cells, kinesin-1 specific transport of cargo from the Golgi appears as trails of fluorescence over time. These findings are further supported by docking studies, which suggest the binding of the activity-based substrate in the nucleotide binding site of kinesin-1.

Date: 2021
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DOI: 10.1038/s41467-021-21626-1

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