State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Biology, College of Chemistry and Chemical Engineering , Hunan University, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province , Changsha 410082 , China.
Anal Chem. 2018 May 15;90(10):6131-6137. doi: 10.1021/acs.analchem.8b00424. Epub 2018 May 2.
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.
细胞表面糖基化包含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以阐明其功能具有重要价值。在这里，我们提出了一种基于杂交链式反应（HCR）的多重荧光共振能量转移（multi-FRET）方法，用于特异性成像细胞表面糖基化。通过在同一糖缀合物上通过代谢糖基化标记安装供体，并通过适体连接的纳米组装体安装受体，由于供体-受体距离接近，会发生分子内 multi-FRET。受益于 HCR 纳米组装体的放大效应和空间灵活性，可以获得特定细胞表面糖基化的增强型 multi-FRET 成像。使用这种基于 HCR 的 multi-FRET 方法，我们在 7211 细胞表面的蛋白质特异性 GalNAcylation 成像中获得了明显的对比度。此外，我们通过可视化 CEM 细胞表面的蛋白质特异性唾液酸化，证明了该方法的通用性。此外，还准确监测了药物处理下 CEM 细胞表面蛋白质特异性唾液酸化的表达变化。这种开发的成像方法可能为研究糖基化功能、发现生物标志物和筛选药物提供有力工具。
