Vesicle Size Regulates Nanotube Formation in the Cell.

Scientific reports (2016-04-08)
Qian Peter Su, Wanqing Du, Qinghua Ji, Boxin Xue, Dong Jiang, Yueyao Zhu, He Ren, Chuanmao Zhang, Jizhong Lou, Li Yu, Yujie Sun
摘要

Intracellular membrane nanotube formation and its dynamics play important roles for cargo transportation and organelle biogenesis. Regarding the regulation mechanisms, while much attention has been paid on the lipid composition and its associated protein molecules, effects of the vesicle size has not been studied in the cell. Giant unilamellar vesicles (GUVs) are often used for in vitro membrane deformation studies, but they are much larger than most intracellular vesicles and the in vitro studies also lack physiological relevance. Here, we use lysosomes and autolysosomes, whose sizes range between 100 nm and 1 μm, as model systems to study the size effects on nanotube formation both in vivo and in vitro. Single molecule observations indicate that driven by kinesin motors, small vesicles (100-200 nm) are mainly transported along the tracks while a remarkable portion of large vesicles (500-1000 nm) form nanotubes. This size effect is further confirmed by in vitro reconstitution assays on liposomes and purified lysosomes and autolysosomes. We also apply Atomic Force Microscopy (AFM) to measure the initiation force for nanotube formation. These results suggest that the size-dependence may be one of the mechanisms for cells to regulate cellular processes involving membrane-deformation, such as the timing of tubulation-mediated vesicle recycling.

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Sigma-Aldrich
链亲和素 来源于阿维丁链霉菌, essentially salt-free, lyophilized powder, ≥13 units/mg protein
Sigma-Aldrich
抗 LAMP1 兔抗, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
溶酶体分离试剂盒, sufficient for 25 g (tissue), sufficient for 20 mL (packed cells), enrichment of lysosomes from tissues and packed cells
Sigma-Aldrich
抗LAMP2 兔抗, affinity isolated antibody, buffered aqueous solution

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