Alkynyl sugar analogs for the labeling and visualization of glycoconjugates in cells
Alkynyl sugar analogs for the labeling and visualization of glycoconjugates in cells
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DOI：
10.1073/pnas.0611307104
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发表时间：
2007-02-20
影响因子：
11.1
通讯作者：
Wong, Chi-Huey
中科院分区：
文献类型：
--
作者：
Hsu, Tsui-Ling;Hanson, Sarah R.;Wong, Chi-Huey
开发用于研究聚糖的细胞活性的工具将有助于描绘病理过程（例如癌症）中异常糖基化的分子基础。将糖报告基团插入细胞糖缀合物中的代谢寡糖工程是一种强大的方法，用于成像聚糖的定位，运输和动力学，并将其分离用于糖蛋白质组学分析。在此，我们表明，炔报告基团可以被纳入细胞聚糖。炔基是小的、惰性的、生物正交柄，其可以通过使用Cu（I）催化的[3 + 2]叠氮化物-炔环加成或点击化学来化学选择性地标记。基于岩藻糖（Fuc）和N-乙酰甘露糖胺（ManNAc）的炔基糖单体被掺入几种癌细胞系中的岩藻糖基化和唾液酸化聚糖中，从而允许糖缀合物的细胞表面和细胞内可视化，以及观察携带炔的糖蛋白。类似于我们以前的结果与叠氮Fuc/炔基探针系统，我们证明了点击激活的荧光探针是有效地和选择性地标记炔基修饰的聚糖的实用工具。由于Fuc和唾液酸是末端聚糖残基，在许多肿瘤中存在显著增加，我们希望我们的方法将提供有关其在癌症中的作用的有用信息，并最终可用于诊断和治疗目的。
Developing tools for investigating the cellular activity of glycans will help to delineate the molecular basis for aberrant glycosylation in pathological processes such as cancer. Metabolic oligosaccharide engineering, which inserts sugar-reporting groups into cellular glycoconjugates, represents a powerful method for imaging the localization, trafficking, and dynamics of glycans and isolating them for glyco-proteomic analysis. Herein, we show that the alkyne-reporting group can be incorporated into cellular glycans. The alkyne group is a small, inert, bio-orthogonal handle that can be chemoselectively labeled by using the Cu(l) catalyzed [3 + 2] azide-alkyne cycloaddition, or click chemistry. Alkynyl sugar monomers, based on fucose (Fuc) and N-acetylmannosamine (ManNAc), were incorporated into fucosylated and sialylated glycans in several cancer cell lines, allowing for cell surface and intracellular visualization of glycoconjugates, as well as, observation of alkyne-bearing glycoproteins. Similarly to our previous results with an azido Fuc/alkynyl probe system, we demonstrated that click-activated fluorogenic probes are practical tools for efficiently and selectively labeling alkynyl-modified glycans. Because Fuc and sialic acid are terminal glycan residues with a notably increased presence in many tumors, we hope that our method will provide useful information about their roles in cancer and ultimately can be used for diagnostic and therapeutic purposes.