Amplified Visualization of Protein-Specific Glycosylation in Zebrafish via Proximity-Induced Hybridization Chain Reaction
Amplified Visualization of Protein-Specific Glycosylation in Zebrafish via Proximity-Induced Hybridization Chain Reaction
复制标题
通过邻近诱导杂交链反应放大斑马鱼蛋白质特异性糖基化的可视化
DOI：
10.1021/jacs.8b08442
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发表时间：
2018-12-05
影响因子：
15
通讯作者：
Yang, Huang-Hao
中科院分区：
文献类型：
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作者：
Li, Jingying;Liu, Shuya;Yang, Huang-Hao
体内特定蛋白质糖基化状态的可视化对于揭示其在疾病发展中的作用具有重要意义。然而，糖基化的普遍存在使得探测某种蛋白质上的聚糖就像大海捞针一样困难。在这里，我们展示了一个接近诱导的杂交链反应（HCR）的策略，蛋白质特异性糖基化的放大可视化。该策略依赖于设计两种DNA探针，聚糖转换探针和蛋白质识别探针，它们分别连接到聚糖和靶蛋白。在与靶序列结合后，两个探针之间发生邻近诱导的杂交，这导致蛋白质识别探针的结构转换，随后触发HCR组装。这种策略已被用于可视化酪氨酸蛋白激酶7特异性唾液酸在活的CEM细胞和斑马鱼，并监测其在药物治疗过程中的变化。它为研究蛋白质特异性糖基化和动态糖链状态与疾病过程的关系提供了一种潜在的工具。
The visualization of glycosylation states of specific proteins in vivo is of great importance for uncovering their roles in disease development. However, the ubiquity of glycosylation makes probing the glycans on a certain protein as difficult as looking for a needle in a haystack. Herein, we demonstrate a proximity-induced hybridization chain reaction (HCR) strategy for amplified visualization of protein-specific glycosylation. The strategy relies on designing two kinds of DNA probes, glycan conversion probes and protein recognition probes, which are attached to glycans and target proteins, respectively. Upon sequential binding to the targets, the proximity-induced hybridization between two probes occurs, which leads to the structure switching of protein recognition probes, followed by triggering of HCR assembly. This strategy has been used to visualize tyrosine-protein kinase 7-specific sialic acid in living CEM cells and zebrafish and to monitor its variation during drug treatment. It provides a potential tool for investigating protein-specific glycosylation and researching the relation between dynamic glycans state and disease process.