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Zeleny, R; Leonard, R; Dorfner, G; Dalik, T; Kolarich, D; Altmann, F.
(2006): Molecular cloning and characterization of a plant alpha1,3/4-fucosidase based on sequence tags from almond fucosidase I.
Phytochemistry. 2006; 67(7):641-648 FullText FullText_BOKU

Abstract:
Out-work with almond peptide N-glycosidase A made us interested also in the alpha 1,3/4-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 alpha 1,2-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 alpha 1,3/4-fucosidase also confirmed the identity of the purified almond enzyme and thus settles the notorious uncertainty about its molecular mass. The alpha 1,3/4-fucosidase from Arabidopsis exhibited striking sequence similarity with an enzyme of similar substrate specificity from Streptomyces sp. (Q9Z4I9) and with putative proteins from rice. (c) 2006 Elsevier Ltd. All rights reserved.
Authors BOKU Wien:
Altmann Friedrich
Dalik Thomas
Leonard Renaud
Find related publications in this database (using NML MeSH Indexing)
Acetylglucosamine - metabolism
Arabidopsis - enzymology
Carbohydrate Sequence - enzymology
Cloning, Molecular - enzymology
DNA, Complementary - analysis
Fucose - metabolism
Hydrolysis - metabolism
Lewis Blood-Group System - metabolism
Molecular Sequence Data - metabolism
Molecular Weight - metabolism
Oryza sativa - enzymology
Pichia - enzymology
Prunus - enzymology
Sequence Alignment - enzymology
Spectrometry, Mass, Electrospray Ionization - enzymology
Streptomyces - enzymology
Substrate Specificity - enzymology
alpha-L-Fucosidase - genetics

Find related publications in this database (Keywords)
arabidopsis
fucosidase
Lewis a
almond
plant glycosidase


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