Epidermal growth factor-like (EGF) repeats are found in numerous extracellular or transmembrane proteins including Notch. EGF repeats containing the appropriate consensus sequences can be modified with two unusual types of glycans: O-fucosylation and O-glucosylation. We have identified the glycosyltransferases that catalyze the addition of the first sugar to these consensus sites: protein O-fucosyltransferase 1 (Pofut1) and protein O-glucosyltransferase (Rumi/Poglut1). Recently, we have demonstrated that Rumi/Poglut1 shows protein O-xylosyltransferase activity as well. Here, we describe how we characterize the enzymatic activity of these enzymes, including preparation of the acceptor substrates, using bacterially expressed EGF repeats. © Springer Science+Business Media New York 2013.
CITATION STYLE
Takeuchi, H., & Haltiwanger, R. S. (2013). Enzymatic analysis of the protein O-Glycosyltransferase, rumi, acting toward epidermal growth factor-like (EGF) repeats. Methods in Molecular Biology, 1022, 119–128. https://doi.org/10.1007/978-1-62703-465-4_10
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