Screening Carbohydrate-Active Enzymes - a Fluorescent Solution

在生命的所有领域，复杂碳水化合物的形成需要大量糖基转移酶(GTS)的协同作用，GTS是催化糖从糖核苷酸供体转移到受体寡糖链上的酶。这种糖基化反应是许多基本生物学过程的中心，例如细胞信号、细胞黏附、癌症发生、人类病原体逃避免疫系统，以及在这项拟议研究的背景下，特别有趣的是，产生复杂和动态的3D结构，即植物细胞壁。与其他具有相似大小和生物学意义的蛋白质家族相比，由于缺乏研究GT底物特异性的分子工具，我们对GT生物学的理解仍然是支离破碎的。现有的GT分析通常是劳力和/或成本密集型的，和/或需要专用设备。我们解决这个问题的方法是设计、合成和应用新型的荧光糖探针，这种探针可以用来识别GT并确定它们的底物特异性。

In all domains of life, the formation of complex carbohydrates requires the concerted action of a multitude of glycosyltransferases (GTs), enzymes that catalyse the transfer of a sugar from a sugar-nucleotide donor onto an acceptor oligosaccharide chain. Such glycosylation reactions are central to many fundamental biological processes, such as cell signalling, cellular adhesion, carcinogenesis, the evasion of the immune system by human pathogens and, of particular interest in the context of this proposed study, the generation of the complex and dynamic 3D architecture that is the plant cell wall. In contrast to other protein families of comparable size and biological significance, our understanding of GT biology remains fragmented, due to a lack of molecular tools for the investigation of GT substrate specificity. Existing GT assays are generally labour- and/or cost-intensive and/or require specialist equipment. Our approach to this problem is through the design, synthesis and application of novel fluorescent sugar probes, which can be used to identify GTs and to determine their substrate specificity.