Accelerated enzymatic galactosylation of N -acetylglucosaminolipids in lipid microdomains

Gavin T. Noble, Faye L. Craven, Josef Voglmeir, Robert Šardzík, Sabine L. Flitsch, Simon J. Webb

    Research output: Contribution to journalArticlepeer-review


    A fluoro-tagged N-acetylglucosamine-capped glycolipid that can form lipid microdomains in fluid phospholipid bilayers has been shown to be enzymatically galactosylated by bovine β(1,4)-galactosyltransferase. MALDI MS, HPLC, and LC-MS revealed that the rate of enzymatic transformation was significantly enhanced by lipid clustering; at a 1% mol/mol loading, clustered glycolipids were galactosylated 9-fold faster than glycolipids dispersed across the bilayer surface. The transformation of the GlcNAc "glycocalyx" into a Gal(β1-4)GlcNAc "glycocalyx" relabeled these vesicles, making them susceptible to agglutination by Erythrina cristagalli lectin (ECL). The kinetic parameters for this transformation revealed a lower apparent K m when the substrate lipids were clustered, which is attributed to multivalent binding to an extended substrate cleft around the active site. These observations may have important implications where soluble enzymes act on substrates embedded within cellular lipid rafts. © 2012 American Chemical Society.
    Original languageEnglish
    Pages (from-to)13010-13017
    Number of pages7
    JournalJournal of the American Chemical Society
    Issue number31
    Publication statusPublished - 8 Aug 2012


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