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The Wave complex is intrinsically inactive.

机译：波复合物本质上无效。

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The Wave proteins activate the Arp2/3 complex at the leading edge of migrating cells. The resulting actin polymerization powers the projection of the plasma membrane in lamellipodia and membrane ruffles. The Wave proteins are always found associated with partner proteins. The canonical Wave complex is a stable complex containing five subunits. Even though it is well admitted that this complex plays an essential regulatory role on Wave function, the mechanisms by which Wave proteins are regulated within the complex are still elusive. Even the constitutive activity or inactivity of the complex is controversial. The major difficulty of these assays resides in the long and difficult purification of the Wave complex by a combination of several chromatography steps, which gives an overall low yield and increases the chance of Wave complex denaturation. Here we report a greatly simplified approach to purify the human Wave complex using a stable cell line expressing a tagged subunit and affinity chromatography. This protocol provided us with sufficient amount of pure Wave complex for functional assays. These assays unambiguously established that the Wave complex in its native conformation is intrinsically inactive, indicating that, like WASP proteins, Wave proteins have a masked C-terminal Arp2/3 binding site at resting state. As a consequence, the Wave complex has to be recruited and activated at the plasma membrane to project migration structures. Importantly, the approach we describe here for multiprotein complex purification is likely applicable to a wide range of human multiprotein complexes.
机译：波蛋白在迁移细胞的前缘激活ARP2 / 3复合物。所得肌动蛋白聚合为层状膜和膜荷叶布中的质膜突出。始终发现波蛋白与合作伙伴蛋白相关。规范波复合物是含有五个亚基的稳定复合物。尽管备受良好的是，这种复杂性在波浪功能上发挥着重要的调节作用，但波浪蛋白在复合物中受到的机制仍然难以捉摸。即使是复杂的组成型活性或不活动也是有争议的。这些测定的主要难度在多种色谱步骤的组合中存在于波复合物的长而难以纯化，这给出了总体低产率并增加了波复位变性的机会。在这里，我们通过表达标记的亚基和亲和层析的稳定细胞系报告了一种大大简化的方法来纯化人波复合物。该协议为我们提供了足够量的纯波复合物，用于功能测定。这些测定明确地确定了其天然构象中的波复合物是本质上无活性的，表明如黄蜂蛋白，波蛋白在静静电状态下具有掩蔽的C末端ARP2 / 3结合位点。因此，必须在质膜处招募和激活波复合物，以项目迁移结构。重要的是，我们在此处描述的多蛋白复合物纯化的方法可能适用于各种人类多蛋白复合物。

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