MINFLUX dissects the unimpeded walking of kinesin-1

Raw MINFLUX data files for single molecule localizations, single molecule stage tracking and kinesin-1 tracking.

Matlab scripts for processing the data and reproducing the results of the publication "MINFLUX dissects the unimpeded walking of kinesin-1" by Wolff and Scheiderer.

Abstract:

We introduce an interferometric MINFLUX microscope that records protein movements with up to 1.7 nm/1 ms spatio-temporal precision. While such precision has so far required attaching disproportionately large beads to the protein, MINFLUX requires the detection of only down to ~20 photons from an ~1-nm-sized fluorophore. Thus, we dissect the stepping of the motor protein kinesin-1 on microtubules at up to physiological ATP concentrations. We uncover rotations of the stalk and the heads of load-free kinesin during stepping; that ATP is taken up with a single head bound to the microtubule; and that ATP hydrolysis occurs when both heads are bound. Our results show that MINFLUX quantifies (sub)millisecond conformational changes of proteins with minimal disturbance.