Comparative characterisation of recombinant invertebrate and vertebrate peptide O-Xylosyltransferases

Andrea Brunner, Daniel Kolarich, Josef Voglmeir, Katharina Paschinger, Iain B H Wilson*

*Corresponding author for this work

Research output: Contribution to journalArticle

18 Citations (Scopus)


Chondroitin and heparan sulphates have key functions in animal development and their synthesis is initiated by the action of UDP-α-D-xylose: proteoglycan core protein β-D-xylosyltransferase (EC cDNAs encoding this enzyme have been previously cloned from mammalian species; this in turn facilitated identification of corresponding Caenorhabditis elegans (sqv-6) and Drosophila melanogaster (oxt) genes. In the present study, we report the expression in Pichia pastoris and subsequent assay using either MALDI-TOF MS or RP-HPLC of recombinant forms of the Caenorhabditis xylosyltransferase SQV-6 and the human xylosyltransferase I, in addition to extending our previous studies on the xylosyltransferase from Drosophila. The enzyme activities were tested with a number of peptide substrates based on portions of the human bikunin, human perlecan and Drosophila syndecan core peptides. Whereas a variant of the latter, containing two Ser-Gly motifs was only modified on one of these motifs, the perlecan peptide with three Ser-Gly motifs could be multiply modified in vitro. Using this substrate, we could for the first time follow, by mass spectrometry, the xylosylation of a peptide with multiple xylosyltransferase acceptor motifs.

Original languageEnglish
Pages (from-to)543-554
Number of pages12
JournalGlycoconjugate Journal
Issue number7-8
Publication statusPublished - Nov 2006


  • Glycosaminoglycan
  • Xylosyltransferase

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