A Homogenous Transcreener® UDP² TR-FRET Glycosyltransferase Assay for High Throughput Screening

Glycosyltransferases (GTs) are catalysts for the transfer of a nucleotide sugar donor to a glycosyl acceptor substrate. More than 200 human glycosyltransferases are known to exist and have diverse metabolic and regulatory roles.

Once glycosyltransferase reactions occur, the nucleotide is often a remaining byproduct. In many cases, glycosyltransferases use UDP sugars as donor substrates. Thus, UDP is a product of the reaction. One method to determine glycosyltransferase enzyme activity is to directly measure the UDP formed by the GT. This provides a universal assay technology to assess the activity of any UDP-producing enzyme in real-time.

Transcreener technology uses a simple but highly effective method that consists of an antibody selective to UDP over a UDP sugar donor and a far-red fluorescent tracer. UDP produced in the reaction competes with the tracer changing the fluorescent properties and providing a readout. The assay is a simple mix-and-read format. Perform your enzyme reaction, add the detection reagent, and measure. The simplicity of the system yields robust results but also makes it extremely amiable to HTS.

Glycosyltransferase inhibitors are often targeted for metabolic disorders. Continuing to build an understanding of GT enzymes is step one. Finding therapeutic methods to reign in overactivity requires selective technologies that can help assess the ability to interact with the target. Robust, HTS-ready assays that are accurate yet flexible provide a powerful tool to accelerate discovery.