Flavin-tag: A Facile Method for Site-Specific Labeling of Proteins with a Flavin Fluorophore,Bioconjugate Chemistry

当前位置： X-MOL 学术 › Bioconjugate Chem. › 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.)

Flavin-tag: A Facile Method for Site-Specific Labeling of Proteins with a Flavin Fluorophore
Bioconjugate Chemistry ( IF 4.0 ) Pub Date : 2021-07-24 , DOI: 10.1021/acs.bioconjchem.1c00306
Yapei Tong ₁ , Misun Lee ₁ , Jeroen Drenth ₁ , Marco W Fraaije ₁

Affiliation

Site-specific protein labeling methods are highly valuable tools for research and applications. We present a new protein labeling method that allows covalent attachment of a chromo- and fluorogenic flavin (FMN) to any targeted protein using a short flavinylation peptide-tag. We show that this peptide can be as short as 7 residues and can be located at the N-terminus, C-terminus, or in internal regions of the target protein. Analogous to kinase-catalyzed phosphorylation, the flavin is covalently attached via a stable phosphothreonyl linkage. The site-specific covalent tethering of FMN is accomplished by using a bacterial flavin transferase. The covalent coupling of FMN was shown to work in Escherichia coli and Saccharomyces cerevisiae cells and could be performed in vitro, rendering the “Flavin-tag” method a powerful tool for the selective decoration of proteins with a biocompatible redox-active fluorescent chromophore.

中文翻译：

黄素标签：一种用黄素荧光团对蛋白质进行位点特异性标记的简便方法

位点特异性蛋白质标记方法是非常有价值的研究和应用工具。我们提出了一种新的蛋白质标记方法，该方法允许使用短黄素化肽标签将发色和荧光黄素 (FMN) 共价连接到任何目标蛋白质。我们表明该肽可以短至 7 个残基，并且可以位于目标蛋白的 N 端、C 端或内部区域。类似于激酶催化的磷酸化，黄素通过稳定的磷酸苏糖酰键共价连接。FMN 的位点特异性共价束缚是通过使用细菌黄素转移酶来完成的。FMN 的共价偶联被证明在大肠杆菌和酿酒酵母细胞中起作用，并且可以在体外进行，使“黄素标签”方法成为使用生物相容性氧化还原活性荧光发色团选择性修饰蛋白质的强大工具。

更新日期：2021-08-19

点击分享查看原文

点击收藏

公开下载

阅读更多本刊最新论文本刊介绍/投稿指南11