Combined Use of Unnatural Amino Acids Enables Dual-Color Super-Resolution Imaging of Proteins via Click Chemistry,ACS Nano

当前位置： X-MOL 学术 › ACS Nano › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Combined Use of Unnatural Amino Acids Enables Dual-Color Super-Resolution Imaging of Proteins via Click Chemistry
ACS Nano ( IF 15.8 ) Pub Date : 2018-12-10 00:00:00 , DOI: 10.1021/acsnano.8b06047
Kim-A. Saal ₁ , Frank Richter ₂ , Peter Rehling ₂ , Silvio O. Rizzoli ₁

Affiliation

Recent advances in optical nanoscopy have brought the imaging resolution to the size of the individual macromolecules, thereby setting stringent requirements for the fluorescent labels. Such requirements are optimally fulfilled by the incorporation of unnatural amino acids (UAAs) in the proteins of interest (POIs), followed by fluorophore conjugation via click chemistry. However, this approach has been limited to single POIs in mammalian cells. Here we solve this problem by incorporating different UAAs in different POIs, which are expressed in independent cell sets. The cells are then fused, thereby combining the different proteins and organelles, and are easily imaged by dual-color super-resolution microscopy. This procedure, which we termed Fuse2Click, is simple, requires only the well-established Amber codon, and allows the use of all previously optimized UAAs and tRNA/RS pairs. This should render it a tool of choice for multicolor click-based imaging.

中文翻译：

结合使用非天然氨基酸可通过点击化学实现蛋白质的双色超高分辨率成像

光学纳米技术的最新进展使成像分辨率达到了单个大分子的大小，从而对荧光标记物提出了严格的要求。通过在目标蛋白质（POI）中掺入非天然氨基酸（UAA），然后通过单击化学。但是，这种方法仅限于哺乳动物细胞中的单个POI。在这里，我们通过在不同的POI中合并不同的UAA（在独立的细胞集中表达）来解决此问题。然后将细胞融合，从而结合不同的蛋白质和细胞器，并通过双色超分辨率显微镜轻松成像。这个过程（我们称为Fuse2Click）很简单，只需要建立良好的Amber密码子，并允许使用所有先前优化的UAA和tRNA / RS对。这应该使其成为多色基于单击的成像的首选工具。

更新日期：2018-12-10

点击分享查看原文

点击收藏

公开下载

阅读更多本刊新发论文本刊介绍/投稿指南