The use of hydrophobic synthetic glycosides as acceptors in glycosyltransferase assays |
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| Authors: | Monica M Palcic Louis D Heerze Michael Pierce Ole Hindsgaul |
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| Institution: | (1) The Department of Food Science, University of Alberta, T6G 2P5 Edmonton, Alberta, Canada;(2) The Department of Anatomy and Cell Biology, University of Miami School of Medicine, 33101 Miami, FL, USA;(3) The Department of Chemistry, University of Alberta, T6G 2G2 Edmonton, Alberta, Canada |
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| Abstract: | A general method is described for the assay of glycosyltransferase activity, which makes use of synthetic glycoside acceptors attached to hydrophobic aglycones. The products formed by incubation of an enzyme with acceptor and radiolabelled sugarnucleotide can then be rapidly (one minute) separated from interfering radioactivity by adsorption on to reverse-phase C-18 cartridges. After aqueous washing, products are easily isolated by elution with methanol. The utility of the method for the assay of  (1–4)galactosyltransferase,  (1–2)fucosyltransferase andN-acetylglucosaminyltransferase I and V is demonstrated. |
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| Keywords: | glycosyltransferase assay synthetic glycosides galactosyltransferase fucosyltransferase N-acetylglucosaminyltransferase |
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