利用重组三脚蛋白枢纽对网格蛋白组装和三聚化的调控。

Regulation of clathrin assembly and trimerization defined using recombinant triskelion hubs.

作者信息

Liu S H, Wong M L, Craik C S, Brodsky F M

机构信息

Department of Pharmacy, School of Pharmacy, University of California, San Francisco 94143-0552, USA.

出版信息

Cell. 1995 Oct 20;83(2):257-67. doi: 10.1016/0092-8674(95)90167-1.

DOI:10.1016/0092-8674(95)90167-1

PMID:7585943

Abstract

Clathrin polymerization into a polyhedral vesicle coat drives receptor sorting at cellular membranes during endocytosis and organelle biogenesis. To study clathrin self-assembly, we expressed the C-terminal third of the clathrin heavy chain in bacteria. The recombinant fragment trimerized, bound clathrin light chains, and morphologically resembled the hub domain of the triskelion-shaped clathrin molecule. Self-assembly of recombinant hubs demonstrated a regulatory role for clathrin light chains and for the distal portions of triskelion legs in clathrin coat formation. Deletion mutagenesis of the hub localized a domain mediating light chain binding and clathrin self-assembly and mapped a transferable trimerization domain. These studies define molecular interactions controlling clathrin self-assembly and establish a recombinant system for future analysis.

摘要

网格蛋白聚合成多面体囊泡衣被，在内吞作用和细胞器生物发生过程中驱动细胞膜上的受体分选。为了研究网格蛋白的自组装，我们在细菌中表达了网格蛋白重链的C端三分之一。重组片段三聚化，结合网格蛋白轻链，并且在形态上类似于三腿蛋白形网格蛋白分子的枢纽结构域。重组枢纽的自组装证明了网格蛋白轻链和三腿蛋白腿部远端部分在网格蛋白衣被形成中的调节作用。枢纽的缺失诱变定位了一个介导轻链结合和网格蛋白自组装的结构域，并绘制了一个可转移的三聚化结构域。这些研究定义了控制网格蛋白自组装的分子相互作用，并建立了一个用于未来分析的重组系统。