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Enzymatic Analysis of the Protein O-Glycosyltransferase, Rumi, Acting Toward Epidermal Growth Factor-Like (EGF) Repeats

 Last Update: 2020-11-26
 Source: Internet
 Author: User

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proteins found body

protein analysis

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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.

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