Du Yifei, Xie Ran, Sun Yuting, Fan Xinqi, Chen Xing
College of Chemistry and Molecular Engineering, Peking University, Beijing, China; Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China.
College of Chemistry and Molecular Engineering, Peking University, Beijing, China.
Methods Enzymol. 2018;598:321-353. doi: 10.1016/bs.mie.2017.06.037. Epub 2017 Oct 31.
Metabolic labeling of glycans with sugar chemical reporters (i.e., unnatural sugars bearing a bioorthogonal group), followed by bioorthogonal reaction with imaging probes or affinity tags, has enabled visualization and proteomic analysis of glycosylation in live cells and in living animals. This two-step metabolic glycan labeling strategy has emerged as a powerful tool for probing glycosylation, but suffers from a lack of cell-type selectivity. Here we describe liposome-assisted bioorthogonal reporter (LABOR), a liposome-assisted format of metabolic glycan labeling that allows for cell-selective and tissue-specific glycan imaging and glycoproteomic profiling. After a brief introduction of the principles and applications of LABOR, we provide detailed protocols for performing LABOR in cell culture and in living mice.
用糖化学报告基团（即带有生物正交基团的非天然糖）对聚糖进行代谢标记，随后与成像探针或亲和标签进行生物正交反应，这使得在活细胞和活体动物中对糖基化进行可视化和蛋白质组学分析成为可能。这种两步代谢聚糖标记策略已成为探测糖基化的有力工具，但缺乏细胞类型选择性。在这里，我们描述了脂质体辅助生物正交报告基团（LABOR），这是一种脂质体辅助的代谢聚糖标记形式，可实现细胞选择性和组织特异性的聚糖成像及糖蛋白质组分析。在简要介绍LABOR的原理和应用后，我们提供了在细胞培养和活体小鼠中进行LABOR的详细方案。
