Multibudded tubules formed by COPII on artificial liposomes.

Multibudded tubules formed by COPII on artificial liposomes.
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DOI:
10.1038/srep00017
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发表时间:
2011
期刊:
影响因子:
4.6
通讯作者:
Schekman, Randy
Schekman, Randy
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Bacia, Kirsten;Futai, Eugene;Prinz, Simone;Meister, Annette;Daum, Sebastian;Glatte, Daniela;Briggs, John A. G.;Schekman, Randy

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包被COPII的小泡在内质网上形成,用于将货物运输到高尔基体。我们使用体外重建来检查COPII支架在重塑脂质双层形状方面的作用。为了避免分离步骤和减少机械操作,使用快速共聚焦荧光和冷冻电子显微镜检查巨大的单层囊泡。COPII显示出对高曲率结构的偏好,但也有足够的灵活性来结合到低曲率。COPII蛋白诱导串珠样收缩的小管,类似于以前在细胞中观察到的。我们推测了从这些多芽的COPII包被的小管中发生囊泡分裂的机械途径,考虑到GTP水解时Sar1两亲性螺旋的退出可能会导致脂质双层的不稳定,从而导致分裂。
COPII-coated vesicles form at the endoplasmic reticulum for cargo transport to the Golgi apparatus. We used in vitro reconstitution to examine the roles of the COPII scaffold in remodeling the shape of a lipid bilayer. Giant Unilamellar Vesicles were examined using fast confocal fluorescence and cryo-electron microscopy in order to avoid separation steps and minimize mechanical manipulation. COPII showed a preference for high curvature structures, but also sufficient flexibility for binding to low curvatures. The COPII proteins induced beads-on-a-string-like constricted tubules, similar to those previously observed in cells. We speculate about a mechanical pathway for vesicle fission from these multibudded COPII-coated tubules, considering the possibility that withdrawal of the Sar1 amphipathic helix upon GTP hydrolysis leads to lipid bilayer destabilization resulting in fission.
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