Journal article
A Direct Fluorescent Activity Assay for Glycosyltransferases Enables Convenient High‐Throughput Screening: Application to O‐GlcNAc Transferase
Matthew G Alteen, Christina Gros, Richard W Meek, David A Cardoso, Jil A Busmann, Gontran Sangouard, Matthew C Deen, Hong‐Yee Tan, David L Shen, Cecilia C Russell, Gideon J Davies, Phillip J Robinson, Adam McCluskey, David J Vocadlo
Angewandte Chemie | Wiley | Published : 2020
Abstract
AbstractGlycosyltransferases carry out important cellular functions in species ranging from bacteria to humans. Despite their essential roles in biology, simple and robust activity assays that can be easily applied to high‐throughput screening for inhibitors of these enzymes have been challenging to develop. Herein, we report a bead‐based strategy to measure the group‐transfer activity of glycosyltransferases sensitively using simple fluorescence measurements, without the need for coupled enzymes or secondary reactions. We validate the performance and accuracy of the assay using O‐GlcNAc transferase (OGT) as a model system through detailed Michaelis–Menten kinetic analysis of various substra..
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Awarded by Natural Sciences and Engineering Research Council of Canada
Awarded by Canadian Glycomics Network
Awarded by National Health and Medical Research Council
Awarded by Australian Research Council