The structural complexity of glycosylation restrains the functional characterization of glycans. We present a versatile carbohydrate ligation technique based on the reaction of cyclic carbamates with primary amines. Cyclic-carbamate-derivatized carbohydrates can be added to primary amine-containing molecules in aqueous solution to yield glycoconjugates. This method enabled the presentation of carbohydrate epitopes on live animal cells, as shown by the acquisition of E-selectin binding sites on mouse MC-38 cells decorated with 3-fucosyllactose or 3-fucosyl-3-sialyllactose. Ligation of 3- and 6-sialyllactose toEscherichia colidemonstrated the importance of sialic acid linkages in regulating complement factor H binding. Proteins were modified with oligosaccharides to study their role in stimulating cytokine secretion by dendritic cells, thus pointing to interactions between glycoproteins and phosphoinositide 3-kinase signaling in controlling interleukin-12, tumor necrosis factor alpha and interleukin-1β release. Overall, cyclic-carbamate-mediated ligation is useful to study the biology of carbohydrate epitopes on proteins and on cell membranes.

中文翻译：

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使用环状氨基甲酸酯衍生的寡糖自定义糖基化的细胞和蛋白质

糖基化的结构复杂性限制了聚糖的功能表征。我们提出了一种基于环状氨基甲酸酯与伯胺反应的通用碳水化合物连接技术。可以将环状氨基甲酸酯衍生的碳水化合物添加到水溶液中的含伯胺的分子中，以产生糖缀合物。这种方法能够在活体动物细胞上呈现碳水化合物表位，如在用3-岩藻糖基乳糖或3-岩藻糖基-3-唾液酸乳糖修饰的小鼠MC-38细胞上获得E-选择蛋白结合位点所示。3-和6-唾液酸乳糖与大肠杆菌的连接证明了唾液酸键在调节补体因子H结合中的重要性。用寡糖修饰蛋白质，以研究其在刺激树突状细胞分泌细胞因子中的作用，因此指出糖蛋白和磷酸肌醇3-激酶信号传导之间的相互作用可控制白介素12，肿瘤坏死因子α和白介素1β的释放。总体而言，环状氨基甲酸酯介导的连接可用于研究蛋白质和细胞膜上碳水化合物表位的生物学特性。