Centre for Mechanochemical Cell Biology, Warwick Medical School, Gibbet Hill Road, Coventry, CV4 7AL, UK.
Nat Commun. 2018 Jul 4;9(1):2604. doi: 10.1038/s41467-018-04993-0.
A current challenge is to develop tags to precisely visualize proteins in cells by light and electron microscopy. Here, we introduce FerriTag, a genetically-encoded chemically-inducible tag for correlative light-electron microscopy. FerriTag is a fluorescent recombinant electron-dense ferritin particle that can be attached to a protein-of-interest using rapamycin-induced heterodimerization. We demonstrate the utility of FerriTag for correlative light-electron microscopy by labeling proteins associated with various intracellular structures including mitochondria, plasma membrane, and clathrin-coated pits and vesicles. FerriTagging has a good signal-to-noise ratio and a labeling resolution of approximately 10 nm. We demonstrate how FerriTagging allows nanoscale mapping of protein location relative to a subcellular structure, and use it to detail the distribution and conformation of huntingtin-interacting protein 1 related (HIP1R) in and around clathrin-coated pits.
目前的一个挑战是开发标签，以便通过光和电子显微镜精确地可视化细胞中的蛋白质。在这里，我们引入 FerriTag，这是一种用于相关光电子显微镜的遗传编码的化学诱导标签。FerriTag 是一种荧光重组电子致密铁蛋白颗粒，可以使用雷帕霉素诱导的异二聚化来连接到感兴趣的蛋白质上。我们通过标记与各种细胞内结构（包括线粒体、质膜和网格蛋白包被的陷窝和小泡）相关的蛋白质，证明了 FerriTag 用于相关光电子显微镜的实用性。FerriTagging 具有良好的信噪比和大约 10nm 的标记分辨率。我们展示了 FerriTagging 如何允许相对于亚细胞结构对蛋白质位置进行纳米尺度的映射，并使用它来详细描述网格蛋白包被的陷窝内外 huntingtin 相互作用蛋白 1 相关（HIP1R）的分布和构象。
