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分子内触发重排诱导的 HCR 用于增强蛋白质特异性糖基化的检测。

Intramolecular trigger remodeling-induced HCR for amplified detection of protein-specific glycosylation.

机构信息

College of Biology, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Key Laboratory for Bio-Nanotechnology and Molecule Engineering of Hunan Province, Changsha, 410082, China.

Department of Pathophysiology, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, Henan, 450001, China.

出版信息

Talanta. 2020 Aug 1;215:120889. doi: 10.1016/j.talanta.2020.120889. Epub 2020 Mar 7.

Abstract

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.

摘要

细胞表面蛋白糖基化的动态变化可作为反映细胞生理状态和疾病发展的重要指标。增强对蛋白特异性糖基化的可视化显示对于解释其功能和机制具有重要价值。因此,我们提出了一种基于分子内触发重构诱导杂交链式反应(HCR)的方法来对蛋白特异性糖基化进行成像。该策略依赖于设计两种 DNA 探针,即蛋白探针和聚糖探针,分别通过适体识别和糖基化通过代谢寡糖工程(MOE)标记在蛋白上。当同一个糖蛋白被标记时,两个探针的互补结构域诱导杂交,从而重构完整的触发物,随后引发 HCR 组装。应用该策略,我们成功地实现了 CEM 细胞表面特异性蛋白糖基化的成像,并监测了药物处理后糖基化的动态变化。它为活细胞中蛋白特异性糖基化的研究领域提供了一种具有高灵活性、特异性和灵敏度的强大工具。

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