Molecular cloning and characterization of a plant α1,3/4-fucosidase based on sequence tags from almond fucosidase I

Reinhard Zeleny, Renaud Leonard, Georg Dorfner, Thomas Dalik, Daniel Kolarich, Friedrich Altmann*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

31 Citations (Scopus)

Abstract

Our work with almond peptide N-glycosidase A made us interested also in the α1,3/4-fucosidase which is used as a specific reagent for glycoconjugate analysis. The enzyme was purified to presumed homogeneity by a series of chromatographic steps including dye affinity and fast-performance anion exchange chromatography. The 63 kDa band was analyzed by tandem mass spectrometry which yielded several partial sequences. A homology search retrieved the hypothetical protein Q8GW72 from Arabidopsis thaliana. This protein has recently been described as being specific for α1,2-linkages. However, cDNA cloning and expression in Pichia pastoris of the A. thaliana fucosidase showed that it hydrolyzed fucose in 3- and 4-linkage to GlcNAc in Lewis determinants whereas neither 2-linked fucose nor fucose in 3-linkage to the innermost GlcNAc residue were attacked. This first cloning of a plant α1,3/4-fucosidase also confirmed the identity of the purified almond enzyme and thus settles the notorious uncertainty about its molecular mass. The α1,3/4-fucosidase from Arabidopsis exhibited striking sequence similarity with an enzyme of similar substrate specificity from Streptomyces sp. (Q9Z4I9) and with putative proteins from rice.

Original languageEnglish
Pages (from-to)641-648
Number of pages8
JournalPhytochemistry
Volume67
Issue number7
DOIs
Publication statusPublished - Apr 2006

Bibliographical note

A corrigendum for this article exists in Phytochemistry, vol. 67, issue 13, p. 1399. DOI: 10.1016/j.phytochem.2006.04.022

Keywords

  • Almond
  • Arabidopsis
  • Fucosidase
  • Lewis a
  • Plant glycosidase

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