Intramolecular trigger remodeling-induced HCR for amplified detection of protein-specific glycosylation
Intramolecular trigger remodeling-induced HCR for amplified detection of protein-specific glycosylation
复制标题
分子内触发重构诱导的 HCR 用于放大检测蛋白质特异性糖基化
DOI：
10.1016/j.talanta.2020.120889
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发表时间：
2020
期刊：
影响因子：
6.1
通讯作者：
Kemin Wang
中科院分区：
文献类型：
--
作者：
Zenghui Li;Baoyin Yuan;Xiaoxia Lin;Xiangxian Meng;Xiaohong Wen;Qiuping Guo;Lie Li;Huishan Jiang;Kemin Wang
细胞表面蛋白质糖基化的动态变化是反映细胞生理状态和疾病发展的重要指标。增强蛋白质特异性糖基化的可视化对于解释其功能和机制具有重要价值。因此，我们提出了一种分子内触发重塑诱导的杂交链反应（HCR）成像蛋白质特异性糖基化。该策略依赖于设计两种DNA探针，蛋白质和聚糖探针，分别通过适体识别标记在蛋白质上和通过代谢寡糖工程（莫伊）标记在聚糖上。在相同的糖蛋白被标记后，两个探针的互补结构域诱导杂交，从而重塑完整的触发物，随后启动HCR组装。应用该方法，我们成功地实现了CEM细胞表面特异性蛋白质糖基化的成像，并监测了药物作用后糖基化的动态变化。它为活细胞上蛋白质特异性糖基化研究领域提供了一种具有高度灵活性、特异性和灵敏度的强有力工具。
Dynamic changes of protein-glycosylation on cell surface act as an important indicator that reflects cellular physiological states and disease developments. The enhanced visualization of protein-specific glycosylation is of great value to interpret its functions and mechanisms. Hence, we present an intramolecular trigger remodeling-induced hybridization chain reaction (HCR) for imaging protein-specific glycosylation. This strategy relies on designing two DNA probes, protein and glycan probes, labeled respectively on protein by aptamer recognition and glycan through metabolic oligosaccharide engineering (MOE). Upon the same glycoprotein was labeled, the complementary domain of two probes induces hybridization and thus to remodel an intact trigger, followed by initiating HCR assembly. Applying this strategy, we successfully achieved imaging of specific protein-glycosylation on CEM cell surface and monitored dynamic changes of the glycosylation after treating with drugs. It provides a powerful tool with high flexibility, specificity and sensitivity in the research field of protein-specific glycosylation on living cells.