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HPLC method for the determination of Fuc to Asn-linked GlcNAc fucosyltransferases

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Abstract

Mammalian cells often contain an enzyme which transfers fucose onto the reducing terminal GlcNAc (GlcNAc-1) of N-glycans with an α1,6-linkage. In plants, on the other hand, the fucose is transferred to GlcNAc-1 with an α1,3-linkage. Insect cells can exhibit both enzymatic activities. Hitherto, the activity of these fucosyltransferases has been determined by the incorporation of radioactively labelled fucose into an acceptor glycopeptide. This assay, however, cannot discriminate these two activities. Here we report on the use of dansylated glycoasparagine for the specific determination of 1,3- and 1,6-fucosyltransferases. The two possible products and the substrate are separated on a reversed phase column and detected by fluorescence.

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Authors and Affiliations

  1. Institut fur Chemie, Universitat fur Bodenkultur, Muthgasse 18, A-1190, Wien, Austria

    Andreas Roitinger, Harald Leiter, Erika Staudacher & Friedrich Altmann

Authors
  1. Andreas Roitinger
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  2. Harald Leiter
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  3. Erika Staudacher
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  4. Friedrich Altmann
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Roitinger, A., Leiter, H., Staudacher, E. et al. HPLC method for the determination of Fuc to Asn-linked GlcNAc fucosyltransferases. Glycoconj J 15, 89–91 (1998). https://doi.org/10.1023/A:1006951802623

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  • DOI: https://doi.org/10.1023/A:1006951802623

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