Arabidopsis thaliana β1,2-xylosyltransferase: an unusual glycosyltransferase with the potential to act at multiple stages of the plant N-glycosylation pathway

Peter Bencúr, Herta Steinkellner, Barbara Svoboda, Jan Mucha, Richard Strasser, Daniel Kolarich, Stephan Hann, Gunda Köllensperger, Josef Glössl, Friedrich Altmann, Lukas Mach*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

51 Citations (Scopus)

Abstract

Xy1T (β1,2-xylosyltransferase) is a unique Golgi-bound glycosyl-transferase that is involved in the biosynthesis of glycoprotein-bound N-glycans in plants. To delineate the catalytic domain of Xy1T, a series of N-terminal deletion mutants was heterologously expressed in insect cells. Whereas the first 54 residues could be deleted without affecting the catalytic activity of the enzyme, removal of an additional five amino acids led to the formation of an inactive protein. Characterization of the N-glycosylation status of recombinant Xy1T revealed that all three potential N-glycosylation sites of the protein are occupied by N-linked oligosaccharides. However, an unglycosylated version of the enzyme displayed substantial catalytic activity, demonstrating that N-glycosylation is not essential for proper folding of Xy1T. In contrast with most other glycosyltransferases, Xy1T is enzymatically active in the absence of added metal ions. This feature is not due to any metal ion directly associated with the enzyme. The precise acceptor substrate specificity of Xy1T was assessed with several physiologically relevant compounds and the xylosylated reaction products were subsequently tested as substrates of other Golgi-resident glycosyltransferases. These experiments revealed that the substrate specificity of Xy1T permits the enzyme to act at multiple stages of the plant N-glycosylation pathway.

Original languageEnglish
Pages (from-to)515-525
Number of pages11
JournalBiochemical Journal
Volume388
Issue number2
DOIs
Publication statusPublished - 1 Jun 2005

Keywords

  • Arabidopsis
  • Glycosyltransferase
  • Golgi apparatus
  • N-glycan biosynthesis
  • Proteolytic processing
  • Xylosyltransferase

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