Precise mapping of subunits in multiprotein complexes by a versatile electron microscopy label
Precise mapping of subunits in multiprotein complexes by a versatile electron microscopy label
复制标题
DOI：
10.1038/nsmb.1811
复制
发表时间：
2010-06-01
影响因子：
16.8
通讯作者：
Hurt, Ed
中科院分区：
文献类型：
--
作者：
Flemming, Dirk;Thierbach, Karsten;Hurt, Ed
多蛋白组装体中亚基的位置知识对于理解其功能至关重要。通过电子显微镜对蛋白质复合物进行拓扑分析已经取得了令人瞩目的发展，但对单个亚基的精确定位的分析却落后了。在这里，我们开发了一种类似于 80 个残基的可克隆标签，在附着到目标蛋白后，可以在体外招募结构突出的电子显微镜标签。该标签在单个颗粒上很容易看到，并且在图像处理和分类后变得异常清晰。因此，我们的方法适用于大分子复合物中亚基的精确拓扑映射。
Positional knowledge of subunits within multiprotein assemblies is crucial for understanding their function. The topological analysis of protein complexes by electron microscopy has undergone impressive development, but analysis of the exact positioning of single subunits has lagged behind. Here we have developed a clonable similar to 80-residue tag that, upon attachment to a target protein, can recruit a structurally prominent electron microscopy label in vitro. This tag is readily visible on single particles and becomes exceptionally distinct after image processing and classification. Thus, our method is applicable for the exact topological mapping of subunits in macromolecular complexes.