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Long‐Range Transport of Giant Vesicles along Microtubule Networks
Author(s) -
Herold Christoph,
Leduc Cécile,
Stock Robert,
Diez Stefan,
Schwille Petra
Publication year - 2012
Publication title -
chemphyschem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.016
H-Index - 140
eISSN - 1439-7641
pISSN - 1439-4235
DOI - 10.1002/cphc.201100669
Subject(s) - microtubule , total internal reflection fluorescence microscope , vesicle , kinesin , vesicular transport protein , molecular motor , motor protein , biophysics , nanobiotechnology , dynein , chemistry , nanotechnology , membrane , biology , microbiology and biotechnology , materials science , biochemistry , nanoparticle
We report on a minimal system to mimic intracellular transport of membrane‐bounded, vesicular cargo. In a cell‐free assay, purified kinesin‐1 motor proteins were directly anchored to the membrane of giant unilamellar vesicles, and their movement studied along two‐dimensional microtubule networks. Motion‐tracking of vesicles with diameters of 1–3 μm revealed traveling distances up to the millimeter range. The transport velocities were identical to velocities of cargo‐free motors. Using total internal reflection fluorescence (TIRF) microscopy, we were able to estimate the number of GFP‐labeled motors involved in the transport of a single vesicle. We found that the vesicles were transported by the cooperative activity of typically 5–10 motor molecules. The presented assay is expected to open up further applications in the field of synthetic biology, aiming at the in vitro reconstitution of sub‐cellular multi‐motor transport systems. It may also find applications in bionanotechnology, where the controlled long‐range transport of artificial cargo is a promising means to advance current lab‐on‐a‐chip systems.