Screening Carbohydrate-Active Enzymes - a Fluorescent Solution

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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.