Abstract
Molecular motors are pivotal for intracellular transport as well as cell motility and have great potential to be put to use outside cells. Here, we exploit engineered motor proteins in combination with self-assembly of actin filaments to actively pull lipid nanotubes from giant unilamellar vesicles (GUVs). In particular, actin filaments are bound to the outer GUV membrane and the GUVs are seeded on a heavy meromyosin-coated substrate. Upon addition of ATP, hollow lipid nanotubes with a length of tens of micrometer are pulled from single GUVs due to the motor activity. We employ the same mechanism to pull lipid nanotubes from different types of cells. We find that the length and number of nanotubes critically depends on the cell type, whereby suspension cells form bigger networks than adherent cells. This suggests that molecular machines can be used to exert forces on living cells to probe membrane-to-cortex attachment.
Highlights
Molecular motors are pivotal for intracellular transport as well as cell motility and have great potential to be put to use outside cells
By tracking individual cells over time, we find that the pulling of lipid nanotubes mediated by motile actin filaments sets in after about 5 min after the attachment of cells to the HMM (Figure 3a)
Since actin-mediated structures provide support of the cell shape and are linked to the cell membrane forming the actin cortex, we focused on the intracellular actin filament organization and dynamics in proximity of lipid nanotubes
Summary
Molecular motors are pivotal for intracellular transport as well as cell motility and have great potential to be put to use outside cells. We quantify the network length per GUV for random and aligned actin filaments and GUVs containing 0 or 20 mol % biotinylated lipids (Figure 2f,g).
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