Biophysical Engineering Group, Max Planck Institute for Medical Research, Jahnstraße 29, D-69120 Heidelberg, Germany.
Department of Physics and Astronomy, Heidelberg University, D-69120 Heidelberg, Germany.
Nano Lett. 2022 Feb 9;22(3):1145-1150. doi: 10.1021/acs.nanolett.1c04254. Epub 2022 Jan 28.
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.
分子马达对于细胞内运输以及细胞运动至关重要,并且在细胞外也有很大的应用潜力。在这里,我们利用工程化的马达蛋白与肌动蛋白丝的自组装相结合,从巨大的单层囊泡(GUV)中主动拉动脂质纳米管。具体来说,肌动蛋白丝与外 GUV 膜结合,并且 GUV 接种在重酶切肌球蛋白涂覆的基底上。加入 ATP 后,由于马达的活性,从单个 GUV 中拉出了数十微米长的中空脂质纳米管。我们使用相同的机制从不同类型的细胞中拉出脂质纳米管。我们发现纳米管的长度和数量与细胞类型密切相关,悬浮细胞形成的网络比贴壁细胞更大。这表明分子机器可用于对活细胞施加力,以探测膜与皮质的连接。