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

doi:10.1016/j.phytochem.2006.01.021

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 journal › Article › peer-review

34 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 language	English
Pages (from-to)	641-648
Number of pages	8
Journal	Phytochemistry
Volume	67
Issue number	7
DOIs	https://doi.org/10.1016/j.phytochem.2006.01.021
Publication status	Published - 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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10.1016/j.phytochem.2006.01.021

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

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.",

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author = "Reinhard Zeleny and Renaud Leonard and Georg Dorfner and Thomas Dalik and Daniel Kolarich and Friedrich Altmann",

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T1 - Molecular cloning and characterization of a plant α1,3/4-fucosidase based on sequence tags from almond fucosidase I

AU - Zeleny, Reinhard

AU - Leonard, Renaud

AU - Dorfner, Georg

AU - Dalik, Thomas

AU - Kolarich, Daniel

AU - Altmann, Friedrich

N1 - A corrigendum for this article exists in Phytochemistry, vol. 67, issue 13, p. 1399. DOI: 10.1016/j.phytochem.2006.04.022

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KW - Fucosidase

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ER -

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

Abstract

Bibliographical note

Keywords

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