网格蛋白包被囊泡内吞过程中AP2α附属枢纽的演变性质
Evolving nature of the AP2 alpha-appendage hub during clathrin-coated vesicle endocytosis.
作者信息
Praefcke Gerrit J K, Ford Marijn G J, Schmid Eva M, Olesen Lene E, Gallop Jennifer L, Peak-Chew Sew-Yeu, Vallis Yvonne, Babu M Madan, Mills Ian G, McMahon Harvey T
机构信息
Medical Research Council Laboratory of Molecular Biology, Cambridge, UK.
出版信息
EMBO J. 2004 Nov 10;23(22):4371-83. doi: 10.1038/sj.emboj.7600445. Epub 2004 Oct 21.
Clathrin-mediated endocytosis involves the assembly of a network of proteins that select cargo, modify membrane shape and drive invagination, vesicle scission and uncoating. This network is initially assembled around adaptor protein (AP) appendage domains, which are protein interaction hubs. Using crystallography, we show that FxDxF and WVxF peptide motifs from synaptojanin bind to distinct subdomains on alpha-appendages, called 'top' and 'side' sites. Appendages use both these sites to interact with their binding partners in vitro and in vivo. Occupation of both sites simultaneously results in high-affinity reversible interactions with lone appendages (e.g. eps15 and epsin1). Proteins with multiple copies of only one type of motif bind multiple appendages and so will aid adaptor clustering. These clustered alpha(appendage)-hubs have altered properties where they can sample many different binding partners, which in turn can interact with each other and indirectly with clathrin. In the final coated vesicle, most appendage binding partners are absent and thus the functional status of the appendage domain as an interaction hub is temporal and transitory giving directionality to vesicle assembly.
网格蛋白介导的内吞作用涉及一组蛋白质网络的组装,这些蛋白质负责选择货物、改变膜形状并驱动内陷、囊泡切割和脱包被。这个网络最初围绕衔接蛋白(AP)附属结构域组装,这些结构域是蛋白质相互作用中心。通过晶体学研究,我们发现突触素的FxDxF和WVxF肽基序与α-附属结构域上不同的亚结构域结合,分别称为“顶部”和“侧面”位点。在体外和体内,附属结构域利用这两个位点与其结合伴侣相互作用。同时占据这两个位点会导致与单个附属结构域(如eps15和epsin1)形成高亲和力的可逆相互作用。仅含有一种基序多个拷贝的蛋白质会结合多个附属结构域,从而有助于衔接蛋白聚集。这些聚集的α(附属结构域)中心具有改变后的特性,它们可以与许多不同的结合伴侣结合,这些结合伴侣又可以相互作用,并间接与网格蛋白相互作用。在最终的包被囊泡中,大多数附属结构域的结合伴侣不存在,因此附属结构域作为相互作用中心的功能状态是暂时的、短暂的,这赋予了囊泡组装方向性。