Lindner R, Ungewickell E
Max-Planck-Institut für Biochemie, Federal Republic of Germany.
Biochemistry. 1991 Sep 17;30(37):9097-101. doi: 10.1021/bi00101a027.
Binding of coated vesicle assembly proteins to clathrin causes it to assemble into regular coat structures. The assembly protein fraction of bovine brain coated vesicles comprises AP180, auxilin, and HA1 and HA2 adaptors. Clathrin heavy chains, separated from their light chains, polymerize with unimpaired efficiency when assembly proteins are added. The reassembled coats were purified by sucrose gradient centrifugation and examined for composition by SDS-PAGE and immunoblotting. We found that all four major coat proteins are incorporated in the presence and absence of light chains. Moreover, each of the purified coat proteins is able to associate directly with clathrin heavy chains in preassembled cages as efficiently as with intact clathrin. We conclude that light chains are not essential for the interaction of AP180, auxilin, and HA1 and HA2 with clathrin.
包被囊泡组装蛋白与网格蛋白的结合使其组装成规则的包被结构。牛脑包被囊泡的组装蛋白部分包括AP180、辅助蛋白以及HA1和HA2衔接蛋白。当加入组装蛋白时，从其轻链分离出的网格蛋白重链能以未受损的效率聚合。通过蔗糖梯度离心法纯化重新组装的包被，并通过SDS - PAGE和免疫印迹法检测其组成。我们发现，无论有无轻链存在，所有四种主要的包被蛋白都能被整合进去。此外，每个纯化的包被蛋白在预组装的笼状结构中与网格蛋白重链直接结合的效率与和完整的网格蛋白结合时一样高。我们得出结论，轻链对于AP180、辅助蛋白以及HA1和HA2与网格蛋白的相互作用并非必不可少。
