Xyloglucan oligosaccharide α-l-fucosidase activity from growing pea stems and germinating nasturtium seeds

Farkas, Vladimir; Hanna, Rami; Maclachlan, Gordon

doi:10.1016/0031-9422(91)83176-l

Cited by 29 publications

(13 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Although there is no previous report on the purification of an afucosidase from plants, a-fucosidase activity (EC 3.2.1.51) has been detected in extracts of almond seed (Kobata, 1982; Ogata-Arakawa et a/., 1977) and etiolated pea stems (O'Neill et a/., 1988). Recently, Farkas and co-workers (Farkas et a/., 1991) reported that the afucosidase activity in pea stems was unaffected by auxin pretreatment and that a-fucosidase activity in germinating nasturtium seeds is developmentally regulated. In the present study, we report on the purification to homogeneity, substrate characterization, and immunogold localization of an a-fucosidase from etiolated pea stems.…”

Section: Introductionmentioning

confidence: 99%

Purification, characterization, and cell wall localization of an α‐fucosidase that inactivates a xyloglucan oligosaccharin

et al. 1993

View full text Add to dashboard Cite

SummaryAn a-fucosidase that releases fucosyl residues from oligosaccharide fragments of xyloglucan, a plant cell wall hemicellulosic polysaccharide, was purified to homogeneity from pea (Pisum satiwum) epicotyls using a combination of cation exchange chromatography and isoelectric focusing. The a-fucosidase has a molecular mass of 20 kDa according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The a-fucosidase has an isoelectric point of 5.5. The substrate specificity of the a-fucosidase was determined by high performance anion exchange chromatographic analysis of oligosaccharide substrates and products. The enzyme hydrolyzes the terminal a-l,2-fucosidic linkage of oligosaccharides and does not cleave pnitrophenyl-a-L-fucoside. The enzyme does not release measurable amounts of fucosyl residues from large polysaccharides. The subcellular localization of a-fucosidase in pea stems and leaves has been studied by immunogold cytochemistry. The a-fucosidase accumulates in primary cell walls and is not detectable in the middle lamella

show abstract

Section: Introductionmentioning

confidence: 99%

Purification, characterization, and cell wall localization of an α‐fucosidase that inactivates a xyloglucan oligosaccharin

et al. 1993

View full text Add to dashboard Cite

show abstract

“…An ␣-fucosidase that removes the ␣-l-fucosyl residue from XXFG has been purified from pea (Pisum sativum) epicotyls (Farkas et al, 1991;Augur et al, 1993) and has been cloned (Augur et al, 1995). Furthermore, cDNAs encoding ␣-l-fucosidase have been also isolated from human (Fukushima et al, 1985;Occhiodoro et al, 1989) and rat (Fisher and Aronson, 1989) livers.…”

mentioning

confidence: 99%

AtFXG1, an Arabidopsis Gene Encoding α-l-Fucosidase Active against Fucosylated Xyloglucan Oligosaccharides

et al. 2002

View full text Add to dashboard Cite

An ␣-l-fucosidase (EC 3.2.1.51) able to release the t-fucosyl residue from the side chain of xyloglucan oligosaccharides has been detected in the leaves of Arabidopsis plants. Moreover, an ␣-l-fucosidase with similar substrate specificity was purified from cabbage (Brassica oleracea) leaves to render a single band on SDS-PAGE. Two peptide sequences were obtained from this protein band, and they were used to identify an Arabidopsis gene coding for an ␣-fucosidase that we propose to call AtFXG1. In addition, an Arabidopsis gene with homology with known ␣-l-fucosidases has been also found, and we proposed to name it as AtFUC1. Both AtFXG1 and ATFUC1 were heterologously expressed in Pichia pastoris cells and the ␣-l-fucosidase activities secreted to the culture medium. The ␣-l-fucosidase encoded by AtFXG1 was active against the oligosaccharides from xyloglucan XXFG as well as against 2Ј-fucosyl-lactitol but not against p-nitrophenyl-␣-lfucopyranoside. However, the AtFUC1 heterologously expressed was active only against 2Ј-fucosyl-lactitol. Thus, the former must be related to xyloglucan metabolism.Plant cell walls are built by two independent networks, cellulose microfibers cross-linked by xyloglucan chains and cross-linked pectins, both networks contributing to their mechanical and functional properties (Roberts, 2001). The xyloglucan-cellulose complex has been considered as the network responsible for controlling the rate of cell expansion, xyloglucan being the load-bearing component in the primary cell walls because of its proposed cross-linking of the cellulose microfibers (Fry, 1989). It consists of a linearattached to the OH-6 of ␤-glucosyl residues. In addition to its structural role, xyloglucan may act as a source of signaling molecules. Oligosaccharides derived from xyloglucan have been found to be formed in vivo (Fry, 1986), and they have been shown to regulate auxin-induced Fry, 1988, 1990) and acid pH-induced (Lorences et al., 1990) growth, the t-Fuc being necessary for the regulatory effect of the XXFG, a nonasaccharide derived from xyloglucan (Fry et al., 1993). The Fuc-deficient mur mutants of Arabidopsis (Reiter et al., 1993) showed dwarf growth habit and fragile cell walls.Enzymes that modify xyloglucan oligosaccharides have been detected in plant cell walls (Fry, 1995). An ␣-fucosidase that removes the ␣-l-fucosyl residue from XXFG has been purified from pea (Pisum sativum) epicotyls (Farkas et al., 1991; Augur et al., 1993) and has been cloned (Augur et al., 1995). Furthermore, cDNAs encoding ␣-l-fucosidase have been also isolated from human (Fukushima et al., 1985;Occhiodoro et al., 1989) and rat (Fisher and Aronson, 1989) livers. However, no ␣-l-fucosidase has yet been identified in Arabidopsis. Although some sequences homologous to pea ␣-l-fucosidase have been found in leguminous plants, there is no clear candidate for this activity in the whole genome of Arabidopsis.Thus, our aim in the present paper has been to look for an Arabidopsis gene(s) encoding for ␣-fucosidase activity. Two differ...

show abstract

“…We could not detect any ␣-fucosidase activity in supernatants or membrane extracts of fruit homogenates that could act on intact fucosylated XG. The assay method that was used readily measured XG nonasaccharide-dependent fucosidase activity in extracts of germinated nasturtium seeds (Farkas et al, 1991).…”

Section: Xg-dependent Fucosyltransferase Activitymentioning

confidence: 99%

“…cellulase. The assay procedure (Farkas et al, 1991) used paper chromatography to measure the initial rate of release of free […”

Section: Enzymic Assaysmentioning

confidence: 99%

“…This would require the action of an ␣-fucosidase with the capacity to defucosylate XG. However, those plant ␣-fucosidases that have been studied to date, those in extracts of germinated nasturtium seeds and pea epicotyls (Farkas et al, 1991;Augur et al, 1993), are only able to hydrolyze Fuc from XG oligosaccharide when it is free in solution, not when it is combined as a subunit in intact XG.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Fucosyltransferase and the Biosynthesis of Storage and Structural Xyloglucan in Developing Nasturtium Fruits

1998

Self Cite

View full text Add to dashboard Cite

Young, developing fruits of nasturtium (Tropaeolum majus L.) accumulate large deposits of nonfucosylated xyloglucan (XG) in periplasmic spaces of cotyledon cells. This "storage" XG can be fucosylated by a nasturtium transferase in vitro, but this does not happen in vivo, even as a transitory signal for secretion. The only XG that is clearly fucosylated in these fruits is the structural fraction (approximately 1% total) that is bound to cellulose in growing primary walls. The two fucosylated subunits that are formed in vitro are identical to those found in structural XG in vivo. The yield of XG-fucosyltransferase activity from membrane fractions is highest per unit fresh weight in the youngest fruits, especially in dissected cotyledons, but declines when storage XG is forming. A block appears to develop in the secretory machinery of young cotyledon cells between sites that galactosylate and those that fucosylate nascent XG. After extensive galactosylation, XG traffic is diverted to the periplasm without fucosylation. The primary walls buried beneath accretions of storage XG eventually swell and lose cohesion, probably because they continue to extend without incorporating components such as fucosylated XG that are needed to maintain wall integrity.

show abstract

Xyloglucan oligosaccharide α-l-fucosidase activity from growing pea stems and germinating nasturtium seeds

Cited by 29 publications

References 28 publications

Purification, characterization, and cell wall localization of an α‐fucosidase that inactivates a xyloglucan oligosaccharin

Purification, characterization, and cell wall localization of an α‐fucosidase that inactivates a xyloglucan oligosaccharin

AtFXG1, an Arabidopsis Gene Encoding α-l-Fucosidase Active against Fucosylated Xyloglucan Oligosaccharides

Fucosyltransferase and the Biosynthesis of Storage and Structural Xyloglucan in Developing Nasturtium Fruits

Contact Info

Product

Resources

About