Udp xylose

Manufactured by Merck Group

UDP-xylose is a chemical compound used in laboratory research. It serves as a substrate for enzymatic reactions and is commonly utilized in the study of carbohydrate metabolism and biosynthesis pathways.

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2 protocols using udp xylose

Enzymatic Characterization of ppGalNAcT

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The enzyme activity of ppGalNAcT and ΔppGalNAcT was determined in 20 μl reaction mixture containing 50 mM MES (2-(N-morpholino)ethanesulfonic acid), pH 7.0, 10 mM MnCl₂, 2 mM UDP-GalNAc (Sigma-Aldrich, Vienna, Austria), 10 nmol acceptor peptide (Cellmano Biotech Co., Ltd., Shanghai, China) and 2 μl enzyme solution (ppGalNAcT or ΔppGalNAcT, protein concentrations 5.0 μg/ml) at 37 °C for 90 min.
For determination of donor specificity UDP-Gal, UDP-GlcNAc, GDP-fucose, UDP-xylose, UDP-glucuronic acid (all from Sigma-Aldrich, Vienna, Austria) and CMP-neuraminic acid (Jennewein Biotechnologie GmBH, Germany) were used instead of UDP-GalNAc. For time courses the standard assay was conducted for 30 h and aliquots were taken at 1, 2, 4, 6, 8, 23, 26 and 30 h. For determination of the position of the transferred GalNAc-residues within the peptide, incubation was carried out with Muc1a, Muc1a’, Muc5Ac and Muc2, respectively (for sequences see Table 1), for 24 h analogously to the standard assay with additions of 10 nmol UDP-GalNAc and 2 μl of enzyme solution after 4, 8 and 20 h.
Each assay was carried out at least in duplicate with appropriate controls.

Taus C., Windwarder M., Altmann F., Grabherr R, & Staudacher E. (2014). UDP-N-acetyl-α-D-galactosamine:polypeptide N-acetylgalactosaminyl-transferase from the snail Biomphalaria glabrata – substrate specificity and preference of glycosylation sites. Glycoconjugate Journal, 31(9), 661-670.

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Synthesis of Nucleotide Sugars

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D-ribose, D-ribulose, and D-ribitol were purchased from Carbosynth. D-galactose was from Merck Millipore. Cytidine triphosphate, D-xylulose, D-xylose, L-arabinose, doxycycline, and 1 mol/L triethylammonium acetate were from Sigma. CMP-N-acetylneuraminic acid, UDPgalactose, UDP-glucose, UDP-N-acetylgalactosamine, UDP-N-acetylglucosamine, GDP-mannose, GDP-fucose, ADP-ribose, UDP-xylose, and UDP-glucuronic acid were from Sigma or Carbosynth.
Rbo5P was produced as previously described (5 ) . CDP-ribitol was synthesized in vitro from Rbo5P and Cytidine triphosphate with recombinantly produced human CRPPA enzyme from Escherichia coli (kind gift of Dr. W. Yue, Oxford) (5 ) . The internal standard, a 13 Cyeast extract containing [U- 13 C]-UDP-glucose, was a kind gift of Dr. G. Hermann, Vienna.

van Tol W., van Scherpenzeel M., Alsady M., Riemersma M., Hermans E., Kragt E., Tasca G., Kamsteeg E.J., Pennings M., van Beusekom E., Vermeulen J.R., van Bokhoven H., Voermans N.C., Willemsen M.A., Ashikov A, & Lefeber D.J. (2019). Cytidine Diphosphate-Ribitol Analysis for Diagnostics and Treatment Monitoring of Cytidine Diphosphate-l-Ribitol Pyrophosphorylase A Muscular Dystrophy. Clinical chemistry, 65(10).

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