Molecular cloning and functional expression of beta1, 2-xylosyltransferase cDNA from Arabidopsis thaliana |
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Authors: | Strasser R Mucha J Mach L Altmann F Wilson I B Glössl J Steinkellner H |
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Affiliation: | Zentrum für Angewandte Genetik, Universit?t für Bodenkultur Wien, Muthgasse 18, A-1190, Vienna, Austria. |
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Abstract: | The transfer of xylose from UDP-xylose to the core beta-linked mannose of N-linked oligosaccharides by beta1,2-xylosyltransferase (XylT) is a widespread feature of plant glycoproteins which renders them immunogenic and allergenic in man. Here, we report the isolation of the Arabidopsis thaliana XylT gene, which contains two introns and encodes a 60.2 kDa protein with a predicted type II transmembrane protein topology typical for Golgi glycosyltransferases. Upon expression of A. thaliana XylT cDNA in the baculovirus/insect cell system, a recombinant protein was produced that exhibited XylT activity in vitro. Furthermore, the recombinant enzyme displayed XylT activity in vivo in the insect cells, as judged by the acquired cross-reaction of cellular glycoproteins with antibodies against the beta1,2-xylose epitope. The cloned XylT cDNA as well as the recombinant enzyme are essential tools to study the role of beta1,2-xylose in the immunogenicity and allergenicity of plant glycoproteins at the molecular level. |
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