Enhanced Imaging of Specific Cell-Surface Glycosylation Based on Multi-FRET.
Enhanced Imaging of Specific Cell-Surface Glycosylation Based on Multi-FRET.
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DOI：
10.1021/acs.analchem.8b00424
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发表时间：
2018-04
影响因子：
7.4
通讯作者：
Baoyin Yuan;Yuanyuan Chen;Yuqiong Sun;Qiuping Guo;Jin Huang;Jianbo Liu;Xiangxian Meng;Xiaohai Yang;Xiaohong Wen;Zenghui Li;Lie Li;Kemin Wang
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文献类型：
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作者：
Baoyin Yuan;Yuanyuan Chen;Yuqiong Sun;Qiuping Guo;Jin Huang;Jianbo Liu;Xiangxian Meng;Xiaohai Yang;Xiaohong Wen;Zenghui Li;Lie Li;Kemin Wang
细胞表面糖基化包含着丰富的反映细胞生理状态的生物学信息，对细胞表面糖基化进行成像研究对阐明其功能具有重要价值。在这里，我们提出了一个杂交链反应（HCR）为基础的多荧光共振能量转移（多FRET）的细胞表面糖基化的特异性成像方法。通过在相同的糖缀合物上通过代谢聚糖标记来安装供体和通过适体拴系的纳米组装体来安装受体，由于供体-受体距离接近而发生分子内多FRET。受益于HCR纳米组装体的放大效应和空间灵活性，可以获得特异性细胞表面糖基化的增强的多FRET成像。利用这种基于HCR的多FRET方法，我们在7211细胞表面蛋白质特异性GalNAc化的成像中实现了明显的对比。此外，我们通过可视化CEM细胞表面上的蛋白质特异性唾液酸化来证明该方法的普遍适用性。此外，准确监测药物处理下CEM细胞表面蛋白特异性唾液酸化的表达变化。该成像方法的发展为研究糖基化功能、发现生物标志物和筛选药物提供了有力的工具。
Cell-surface glycosylation contains abundant biological information that reflects cell physiological state, and it is of great value to image cell-surface glycosylation to elucidate its functions. Here we present a hybridization chain reaction (HCR)-based multifluorescence resonance energy transfer (multi-FRET) method for specific imaging of cell-surface glycosylation. By installing donors through metabolic glycan labeling and acceptors through aptamer-tethered nanoassemblies on the same glycoconjugate, intramolecular multi-FRET occurs due to near donor-acceptor distance. Benefiting from amplified effect and spatial flexibility of the HCR nanoassemblies, enhanced multi-FRET imaging of specific cell-surface glycosylation can be obtained. With this HCR-based multi-FRET method, we achieved obvious contrast in imaging of protein-specific GalNAcylation on 7211 cell surfaces. In addition, we demonstrated the general applicability of this method by visualizing the protein-specific sialylation on CEM cell surfaces. Furthermore, the expression changes of CEM cell-surface protein-specific sialylation under drug treatment was accurately monitored. This developed imaging method may provide a powerful tool in researching glycosylation functions, discovering biomarkers, and screening drugs.