Purification and architecture of the ubiquitous Wave complex
Purification and architecture of the ubiquitous Wave complex
复制标题
DOI：
10.1073/pnas.0400628101
复制
发表时间：
2004-03-30
影响因子：
11.1
通讯作者：
Kirschner, MW
中科院分区：
文献类型：
--
作者：
Gautreau, A;Ho, HYH;Kirschner, MW
Wave蛋白是Arp 2/3复合物的主要激活剂。普遍存在的Wave-2是迁移细胞前沿肌动蛋白聚合所必需的。在这里，我们从HeLa细胞中纯化Wave-2。共纯化了Sra、Nap、Wave-2、Abi和Hspc五种蛋白质，表明它们形成紧密复合物。这些蛋白质仅以复合形式存在，但Hspc除外，其显示游离池。我们重建波-2复杂的共翻译在体外的五个亚基，并使用该系统与特定的免疫沉淀研究复合物的分子结构。该综合体是围绕着Nap和Abi的核心组织的。Sra是在Nap侧募集的外周亚基，而Wave和Hspc亚基在核心的Abi侧募集。
The Wave proteins are major activators of the Arp2/3 complex. The ubiquitous Wave-2 is required for actin polymerization at the leading edge of migrating cells. Here we purify Wave-2 from HeLa cells. Five proteins, Sra, Nap, Wave-2, Abi, and Hspc, are copurified, indicating that they form a tight complex. These proteins are only present in the complexed form, with the exception of Hspc, which displays a free pool. We reconstitute the Wave-2 complex by cotranslating in vitro the five subunits and use this system together with specific immunoprecipitations to study the molecular architecture of the complex. The complex is organized around a core of Nap and Abi. Sra is a peripheral subunit recruited on the Nap side, whereas the Wave and Hspc subunits are recruited on the Abi side of the core.