Probing mucin-type O-linked glycosylation in living animals
Probing mucin-type O-linked glycosylation in living animals
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DOI：
10.1073/pnas.0506855103
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发表时间：
2006-03-28
影响因子：
11.1
通讯作者：
Bertozzi, CR
中科院分区：
文献类型：
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作者：
Dube, DH;Prescher, JA;Bertozzi, CR
已知O-连接蛋白糖基化的变化与疾病状态相关，但由于缺乏实验工具，难以在生理环境中监测。在这里，我们报告了一种技术，用于快速分析O-连接的糖蛋白在活体动物的代谢标记N-叠氮基乙酰半乳糖胺（GalNAz），然后由施陶丁格连接膦探针。在用全乙酰化形式的GalNAz注射小鼠后，在各种组织中观察到叠氮化物标记的糖蛋白，包括肝、肾和心脏、血清和分离的脾细胞。B细胞糖蛋白被GalNAz强烈标记，但T细胞糖蛋白没有，表明糖基化机制或代谢的根本差异。此外，GalNAz标记的B细胞可以选择性地与膦探针通过施陶丁格连接在活的动物。用GalNAz进行代谢标记，然后进行Staudinger连接，为这种翻译后修饰的蛋白质组学分析和鉴定与疾病相关的O-连接糖蛋白指纹提供了一种手段。
Changes in O-linked protein glycosylation are known to correlate with disease states but are difficult to monitor in a physiological setting because of a lack of experimental tools. Here, we report a technique for rapid profiling of O-linked glycoproteins in living animals by metabolic labeling with N-azidoacetylgalactosamine (GalNAz) followed by Staudinger ligation with phosphine probes. After injection of mice with a peracetylated form of GalNAz, azide-labeled glycoproteins were observed in a variety of tissues, including liver, kidney, and heart, in serum, and on isolated splenocytes. B cell glycoproteins were robustly labeled with GalNAz but T cell glycoproteins were not, suggesting fundamental differences in glycosylation machinery or metabolism. Furthermore, GalNAz-labeled B cells could be selectively targeted with a phosphine probe by Staudinger ligation within the living animal. Metabolic labeling with GalNAz followed by Staudinger ligation provides a means for proteomic analysis of this posttranslational modification and for identifying O-linked glycoprotein fingerprints associated with disease.