Molecular cloning and characterization of a plant alpha1,3/4-fucosidase based on sequence tags from almond fucosidase I

Phytochemistry
Reinhard ZelenyF Altmann

Abstract

Our work with almond peptide N-glycosidase A made us interested also in the alpha1,3/4-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 alpha1,2-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 alpha1,3/4-fucosidase also confirmed the identity of the purified almond enzyme and thus settles the notorious uncertainty about its molecular mass. The alpha1,3/4-fucosidase from Arabidopsis exhibited striking sequence similarity with an enzyme of similar substrate specificity from Streptomyces sp. (Q9Z4I9) and with putative proteins from rice.

Citations

Mar 28, 2012·The Journal of Biological Chemistry·Haruko SakuramaTakane Katayama
May 31, 2014·Journal of Agricultural and Food Chemistry·Rosanna ZivoliMichele Solfrizzo
Aug 28, 2015·Bioscience, Biotechnology, and Biochemistry·Md Ziaur RahmanYoshinobu Kimura
Feb 10, 2009·Phytochemistry·Renaud LéonardFriedrich Altmann
Jun 12, 2009·Carbohydrate Research·Gerard J A RouwendalDirk Bosch
Aug 21, 2013·Journal of Experimental Botany·Lenka Franková, Stephen C Fry
Jun 27, 2014·Frontiers in Plant Science·Eva KnochNaomi Geshi
Feb 2, 2019·Analytical and Bioanalytical Chemistry·Tangrong LiuZhe Xu
Apr 26, 2020·Cellular and Molecular Life Sciences : CMLS·Haiyang WuNathalie Juge
Aug 3, 2014·The Journal of Biological Chemistry·Hongnan CaoGeorge N Phillips
Mar 9, 2021·Frontiers in Plant Science·Richard StrasserJosé M Estevez
Jun 21, 2020·Insect Biochemistry and Molecular Biology·Shuntaro NakamuraEnoch Y Park

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