Reconstitution of clathrin-coated bud and vesicle formation with minimal components
Reconstitution of clathrin-coated bud and vesicle formation with minimal components
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DOI:
10.1038/ncb2478
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发表时间:
2012-06-01
影响因子:
21.3
通讯作者:
Ungewickell, Ernst J.
中科院分区:
文献类型:
--
作者:
Dannhauser, Philip N.;Ungewickell, Ernst J.
During the process of clathrin-mediated endocytosis an essentially planar area of membrane has to undergo a gross deformation to form a spherical bud. Three ways have been recognized by which membranes can be induced to transform themselves locally from a planar state to one of high curvature: a change in lipid distribution between the leaflets, insertion of a protein into one leaflet and formation of a protein scaffold over the surface(1). Such a scaffold is spontaneously generated by clathrin(2-5). Conjectures that the attachment of clathrin was the cause of the change in curvature were challenged on theoretical grounds(6), and also by the discovery of a number of clathrin-associated proteins with the capacity to induce membrane curvature(7-9). We have now developed a cell-free system that has enabled us to demonstrate that clathrin polymerization alone is sufficient to generate spherical buds in a membrane. This process is reversible, as shown by the reassimilation of the buds into the planar membrane when the intra-clathrin contacts are dissociated by the chaperone Hsc70. We further show that the final step in the formation of coated vesicles ensues when clathrin-coated buds are released through the action of dynamin.