Alkynyl sugar analogs for the labeling and visualization of glycoconjugates in cells
申请人:WONG CHI-HUEY
公开号:US20080268468A1
公开(公告)日:2008-10-30
The present disclosure relates to a method for metabolic oligosaccharide engineering that incorporates derivatized alkyne-bearing sugar analogs as “tags” into cellular glycoconjugates. The disclosed method incorporates alkynyl derivatized Fuc and alkynyl derivatized ManNAc sugars into a cellular glycoconjugate. A chemical probe comprising an azide group and a visual probe or a fluorogenic probe is used to label the alkyne-derivatized sugar-tagged glycoconjugate. In one aspect, the chemical probe binds covalently to the alkynyl group by Cu(I)-catalyzed [3+2] azide-alkyne cycloaddition and is visualized at the cell surface, intracellularly, or in a cellular extract. The labeled glycoconjugate is capable of detection by flow cytometry, SDS-PAGE, Western blot, ELISA or confocal microscopy, and mass spectrometry.
本公开涉及一种代谢寡糖工程的方法,该方法将衍生化的烷基糖类似物作为“标记”并纳入细胞糖蛋白中。所披露的方法将烷基化的Fuc和烷基化的ManNAc糖纳入细胞糖蛋白中。使用含有偶氮基团的化学探针和视觉探针或荧光探针来标记烷基化糖标记的糖蛋白。在一个方面,化学探针通过Cu(I)-催化的[3+2]偶氮-烷基环加成与烷基基团共价结合,并在细胞表面、细胞内或细胞提取物中可视化。标记的糖蛋白可通过流式细胞术、SDS-PAGE、Western blot、ELISA或共聚焦显微镜和质谱检测。