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Labeling glycans on living cells by a chemoenzymatic glycoengineering approach

机译:通过化学酶促糖工程方法在活细胞上标记聚糖

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Structural glycobiology has traditionally been a challenging field due to a limited set of tools available to investigate the diverse and complex glycan molecules. However, we cannot ignore that glycans play critical roles in health as well as in disease, and are present in more than 50% of all proteins and on over 80% of all surface proteins. Chemoenzymatic glycoengineering (CGE) methods are a powerful set of tools to synthesize complex glycans, but the full potential of these methods have not been explored in cell biology yet. Herein, we report the labeling of live Chinese hamster ovary (CHO) cells by employing three highly specific glycosyltransferases: a sialyltransferase, a galactosyltransferase, and an?N-acetyl-glucosaminyl transferase. We verified our results by bio-orthogonal blots and further rationalized them by computational modeling. We expect CGE applications in cell biology to rise and their implementation will assist in structural-functional discoveries in glycobiology. This research will contribute to this effort.
机译:由于可用于研究多样化和复杂聚糖分子的工具集有限,结构糖生物学一直以来都是具有挑战性的领域。但是,我们不能忽略聚糖在健康和疾病中都起着至关重要的作用,并且存在于所有蛋白质的50%以上和所有表面蛋白质的80%以上。化学酶糖工程(CGE)方法是合成复杂聚糖的功能强大的工具集,但是这些方法的全部潜力尚未在细胞生物学中进行探索。本文中,我们报告了通过使用三种高度特异性的糖基转移酶标记的中国仓鼠卵巢(CHO)细胞标记:唾液酸转移酶,半乳糖基转移酶和?N-乙酰基-氨基葡萄糖氨基转移酶。我们通过生物正交印迹验证了我们的结果,并通过计算模型进一步合理化了结果。我们预计细胞生物学中CGE的应用将增加,其实施将有助于糖生物学的结构功能发现。这项研究将为这项工作做出贡献。

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