Despite the progress in refining static structures of proteins including protein motors the understanding of their chemical and mechanical functions remain indistinct. In this project we advance the accuracy and speed in direct tracking of single protein motion and apply the new method to dissect the mechanisms of directional and diffusible motion of proteins along microtubules. The methodology used in this project is based on recent concept of interferometric imaging of light scattered on nanoparticles or unlabeled proteins (iSCAT). Our preliminary results indicate, that the method is capable of tracking extremely small scattering labels with a nanometer precision at microseconds exposure time. Given this technological leap we advance with new protein labeling strategies employing only a few nanometer long anchors for high fidelity tracking. The combined effect of the proposed methodology will allow us to closely track not only the stepping mechanics of molecular motors but also map the intermediate states driven by the energetic landscapes of the interaction.
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