A Single Excitation‐Duplexed Imaging Strategy for Profiling Cell Surface Protein‐Specific Glycoforms,Angewandte Chemie International Edition

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A Single Excitation‐Duplexed Imaging Strategy for Profiling Cell Surface Protein‐Specific Glycoforms
Angewandte Chemie International Edition ( IF 16.1 ) Pub Date : 2016-03-22 , DOI: 10.1002/anie.201601233
Na Wu ₁ , Lei Bao ₁ , Lin Ding ₁ , Huangxian Ju ₁

Affiliation

This work develops a site‐specific duplexed luminescence resonance energy transfer system on cell surface for simultaneous imaging of two kinds of monosaccharides on a specific protein by single near‐infrared excitation. The single excitation‐duplexed imaging system utilizes aptamer modified upconversion luminescent nanoparticles as an energy donor to target the protein, and two fluorescent dye acceptors to tag two kinds of cell surface monosaccharides by a dual metabolic labeling technique. Upon excitation at 980 nm, only the dyes linked to protein‐specific glycans can be lit up by the donor by two parallel energy transfer processes, for in situ duplexed imaging of glycoforms on specific protein. Using MUC1 as the model, this strategy can visualize distinct glycoforms of MUC1 on various cell types and quantitatively track terminal monosaccharide pattern. This approach provides a versatile platform for profiling protein‐specific glycoforms, thus contributing to the study of the regulation mechanisms of protein functions by glycosylation.

中文翻译：

用于分析细胞表面蛋白特定糖型的单一激发-双重成像策略

这项工作开发了一种在细胞表面上的定点双工发光共振能量转移系统，可通过单次近红外激发同时成像特定蛋白质上的两种单糖。单激发双工成像系统利用适体修饰的上转换发光纳米粒子作为靶向蛋白质的能量供体，并利用两个荧光染料受体通过双重代谢标记技术标记两种细胞表面单糖。在980 nm激发时，供体通过两个平行的能量转移过程仅能点亮与蛋白特异性聚糖连接的染料，以便对特定蛋白上的糖形进行双工成像。使用MUC1作为模型，该策略可以在各种细胞类型上可视化MUC1的独特糖型，并定量跟踪末端单糖模式。该方法提供了一个用于分析蛋白质特异性糖型的通用平台，从而有助于研究糖基化对蛋白质功能的调节机制。

更新日期：2016-03-22

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