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Related Experiment Video

Updated: Jan 29, 2026

Assembling Molecular Shuttles Powered by Reversibly Attached Kinesins

Assembling Molecular Shuttles Powered by Reversibly Attached Kinesins

Published on: January 26, 2019

Assembling Molecular Shuttles Powered by Reversibly Attached Kinesins.

Neda M Bassir Kazeruni¹, Stanislav Tsitkov¹, Henry Hess²

¹Department of Biomedical Engineering, Columbia University.

Journal of Visualized Experiments : Jove

|February 12, 2019

Summary

This study presents a novel method for creating dynamic molecular shuttles using kinesin motor proteins and microtubules. These shuttles exhibit continuous assembly and disassembly, leading to unique dynamic behaviors on surfaces.

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Isolation and Purification of Kinesin from Drosophila Embryos

Isolation and Purification of Kinesin from Drosophila Embryos

Published on: April 27, 2012

Area of Science:

Biophysics
Nanotechnology
Materials Science

Background:

Kinesin motor proteins are crucial for intracellular transport, moving cargo along microtubule tracks.
Existing methods for surface-based kinesin systems often involve strong, irreversible attachments, limiting dynamic behavior.
Developing systems with reversible kinesin-surface interactions is key to creating dynamic molecular machines.

Purpose of the Study:

To describe a protocol for constructing kinesin-powered molecular shuttles with weak and reversible kinesin-surface attachment.
To demonstrate a system where microtubules actively recruit kinesin motors from solution onto a surface.
To investigate the dynamic behaviors arising from the continuous assembly and disassembly of kinesin-microtubule interactions.

Main Methods:

UV-Vis spectrophotometry for reagent concentration determination.
Surface preparation including ozone/UV treatment and silanization of coverslips for flow cell construction.
Total internal reflection fluorescence (TIRF) microscopy for simultaneous imaging of kinesin motors and microtubules.

Main Results:

Successful creation of molecular shuttles with dynamic kinesin-microtubule interactions.
Observation of microtubules recruiting kinesin motors from solution to the surface.
Demonstration of continuous kinesin-microtubule gliding, desorption, and re-recruitment.
Formation of transient kinesin trails due to gliding microtubules.

Conclusions:

This protocol enables the creation of novel, dynamic kinesin-powered molecular shuttles.
The reversible attachment mechanism allows for continuous cycling of motor proteins, leading to emergent dynamic behaviors.
The system provides a platform for studying active matter and designing responsive nanomaterials.