Molecular cloning and functional expression of β1,2‐xylosyltransferase cDNA from Arabidopsis thaliana 1

FEBS Letters

Mucha, J.²  Steinkellner, H.²  Strasser, R.²  Altmann, F.¹  Glössl, J.²  Mach, L.²  Wilson, I.B.H.¹ 

[1] Institut für Chemie, Universität für Bodenkultur Wien, Muthgasse 18, A-1190 Vienna, Austria;Zentrum für Angewandte Genetik, Universität für Bodenkultur Wien, Muthgasse 18, A-1190 Vienna, Austria

关键词: β1;    2-Xylosyltransferase;    Glycosyltransferase;    N-Glycan;    Arabidopsis thaliana;   

DOI  :  10.1016/S0014-5793(00)01443-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The transfer of xylose from UDP-xylose to the core β-linked mannose of N-linked oligosaccharides by β1,2-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 β1,2-xylose epitope. The cloned XylT cDNA as well as the recombinant enzyme are essential tools to study the role of β1,2-xylose in the immunogenicity and allergenicity of plant glycoproteins at the molecular level.

【 授权许可】

Unknown