Berlin 2008 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 12: Cellular Force Generation
BP 12.3: Vortrag
Dienstag, 26. Februar 2008, 18:00–18:15, C 243
Transport of micrometer-sized vesicles by kinesin in vitro — •Christoph Herold1, Cécile Leduc2, Eugene P. Petrov1, Stefan Diez2, and Petra Schwille1 — 1Biophysics / BIOTEC, TU Dresden, Tatzberg 47-51, 01307 Dresden — 2Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstr.
Cytoskeletal motor proteins (e.g., kinesin) are responsible for directed transport in cells. Motor proteins can also be used in artificial bionanotechnological systems to provide a controlled cargo transport. We explore this possibility by using giant unilamellar vesicles (GUVs) as a micrometer-sized cargo model and establish an in vitro system to transport this cargo by kinesin (rK430) molecules along surface-attached microtubules (MTs). Kinesin was linked to GUVs (diameter 1−4 µ m) via biotin−streptavidin interaction. MTs and moving GUVs were visualized using fluorescence wide-field imaging microscopy. We observe directed transport of GUVs along MTs with traveling distances of up to 100 µ m and velocities of ∼ 0.7 µ m/s being in a good agreement with the velocity of kinesin motion along MTs (∼ 0.8 µ m/s). The long walking distances, as well as the visualization of the GFP-labeled kinesin molecules by total internal reflection fluorescence imaging, suggest that a large number ( 10) of kinesin molecules is involved in the transport of a single GUV. Apart from its biotechnological importance, this system might additionally be useful to gain further understanding of vesicle transport processes in cells.