First Isolation of Human UDP-d-Xylose: Proteoglycan Core Protein β-d-Xylosyltransferase Secreted from Cultured JAR Choriocarcinoma Cells

Kühn, Joachim; Götting, Christian; Schnölzer, Martina; Kempf, Tore; Brinkmann, Thomas; Kleesiek, Knut

doi:10.1074/jbc.m005111200

Cited by 70 publications

(58 citation statements)

References 38 publications

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“…Plants, many animals, fungi, Bacteria, and Archaea produce enzymes (UXS) that convert UDP-GlcA to UDP-Xyl (27,(37)(38)(39)(40), whereas UAS forms both UDP-Api and UDP-Xyl in a ratio of ϳ2:1. It is not known whether UXS or UAS is the ancestral gene.…”

Section: Discussionmentioning

confidence: 99%

Functional Characterization of UDP-apiose Synthases from Bryophytes and Green Algae Provides Insight into the Appearance of Apiose-containing Glycans during Plant Evolution

Smith

Yang

Levy³

et al. 2016

Journal of Biological Chemistry

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Apiose is a branched monosaccharide that is present in the cell wall pectic polysaccharides rhamnogalacturonan II and apiogalacturonan and in numerous plant secondary metabolites. These apiose-containing glycans are synthesized using UDPapiose as the donor. UDP-apiose (UDP-Api) together with UDPxylose is formed from UDP-glucuronic acid (UDP-GlcA) by UDP-Api synthase (UAS). It was hypothesized that the ability to form Api distinguishes vascular plants from the avascular plants and green algae. UAS from several dicotyledonous plants has been characterized; however, it is not known if avascular plants or green algae produce this enzyme. Here we report the identification and functional characterization of UAS homologs from avascular plants (mosses, liverwort, and hornwort), from streptophyte green algae, and from a monocot (duckweed). The recombinant UAS homologs all form UDP-Api from UDP-glucuronic acid albeit in different amounts. Apiose was detected in aqueous methanolic extracts of these plants. Apiose was detected in duckweed cell walls but not in the walls of the avascular plants and algae. Overexpressing duckweed UAS in the moss Physcomitrella patens led to an increase in the amounts of aqueous methanol-acetonitrile-soluble apiose but did not result in discernible amounts of cell wall-associated apiose. Thus, bryophytes and algae likely lack the glycosyltransferase machinery required to synthesize apiose-containing cell wall glycans. Nevertheless, these plants may have the ability to form apiosylated secondary metabolites. Our data are the first to provide evidence that the ability to form apiose existed prior to the appearance of rhamnogalacturonan II and apiogalacturonan and provide new insights into the evolution of apiose-containing glycans.

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Section: Discussionmentioning

confidence: 99%

Functional Characterization of UDP-apiose Synthases from Bryophytes and Green Algae Provides Insight into the Appearance of Apiose-containing Glycans during Plant Evolution

Smith

Yang

Levy³

et al. 2016

Journal of Biological Chemistry

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“…Because XylT precedes galactosyltransferase in the pathway, it is reasonable to assume that the association may occur in a more proximal biosynthetic compartment. The recent purification and cloning of XylT should help clarify this issue (33,34).…”

Section: Discussionmentioning

confidence: 99%

Enzyme interactions in heparan sulfate biosynthesis: Uronosyl 5-epimerase and 2- O -sulfotransferase interact in vivo

Pinhal

Smith²,

Olson³

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

142

109

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The formation of heparan sulfate occurs within the lumen of the endoplasmic reticulum-Golgi complex-trans-Golgi network by the concerted action of several glycosyltransferases, an epimerase, and multiple sulfotransferases. In this report, we have examined the location and interaction of tagged forms of five of the biosynthetic enzymes: galactosyltransferase I and glucuronosyltransferase I, required for the formation of the linkage region, and GlcNAc N-deacetylase͞N-sulfotransferase 1, uronosyl 5-epimerase, and uronosyl 2-O-sulfotransferase, the first three enzymes involved in the modification of the chains. All of the enzymes colocalized with the medial-Golgi marker ␣-mannosidase II. To study whether any of these enzymes interacted with each other, they were relocated to the endoplasmic reticulum (ER) by replacing their cytoplasmic N-terminal tails with an ER retention signal derived from the cytoplasmic domain of human invariant chain (p33). Relocating either galactosyltransferase I or glucuronosyltransferase I had no effect on the other's location or activity. However, relocating the epimerase to the ER caused a parallel redistribution of the 2-Osulfotransferase. Transfected epimerase was also located in the ER in a cell mutant lacking the 2-O-sulfotransferase, but moved to the Golgi when the cells were transfected with 2-O-sulfotransferase cDNA. Epimerase activity was depressed in the mutant, but increased upon restoration of 2-O-sulfotransferase, suggesting that their physical association was required for both epimerase stability and translocation to the Golgi. These findings provide in vivo evidence for the formation of complexes among enzymes involved in heparan sulfate biosynthesis. The functional significance of these complexes may relate to the rapidity of heparan sulfate formation.glycosaminoglycans ͉ sulfation ͉ epimerization ͉ enzyme localization ͉ Golgi complex T he biosynthesis of heparan sulfate initiates by the translation of a proteoglycan core protein and the assembly of the so-called linkage region tetrasaccharide on specific serine residues (-GlcA␤1,3Gal␤1,3Gal␤1,4Xyl␤1-O-Ser). The chain then polymerizes by the addition of alternating N-acetylglucosamine (GlcNAc␣1,4) and glucuronic acid (GlcA␤1,4) residues. A series of modification reactions takes place simultaneously that involves at least six enzymatic activities: (i) N-deacetylation of a portion of GlcNAc residues, (ii) N-sulfation of the resulting unsubstituted amino groups to form GlcNS units, (iii) 5-epimerization of adjacent D-GlcA residues to form L-iduronic acid (IdoA), (iv) 2-O-sulfation of L-IdoA and more rarely D-GlcA residues, (v) 6-O-sulfation of glucosamine units, and (vi) occasional 3-O-sulfation of glucosamine residues (Fig. 1A). A major question concerns how these enzymes orchestrate the formation of specific oligosaccharide sequences with unique binding properties for ligands (1, 2). Multiple isozymes exist for several of the transferases that differ in substrate specificity and temporal͞spatial expression during developmen...

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“…Five glycosyltransferases, i.e. EXT1, EXT2, EXTL1, EXTL2, and EXTL3, have been cloned and identified to participate in the synthesis of heparan in addition to the four enzymes for the synthesis of linkage tetrasaccharide (5,6,(15)(16)(17). EXT1 and EXT2 are the heparan synthases that have both ␣1,4-N-acetylglucosaminyltransferase (␣4Gn-T) and ␤1,4-glucuronyltransferase (␤4GlcA-T) activities and elongate the heparan units for polymerization (35)(36)(37).…”

Section: Fig 6 Enzymatic Synthesis Of Chondroitin Pentasaccharidementioning

confidence: 99%

“…The glycosyltransferases, such as glucuronyltransferase-I (GlcAT-I), ␤1,3-galactosyltransferase-VI (␤3Gal-T6), ␤4Gal-T7, and xylosyltransferase, for the linkage have been identified (5,6,(15)(16)(17). Recently, chondroitin synthase (CSS) has been cloned and identified (18).…”

mentioning

confidence: 99%

Enzymatic Synthesis of Chondroitin with a Novel Chondroitin Sulfate N-Acetylgalactosaminyltransferase That Transfers N-Acetylgalactosamine to Glucuronic Acid in Initiation and Elongation of Chondroitin Sulfate Synthesis

Gotoh¹,

Sato²,

Akashima³

et al. 2002

Journal of Biological Chemistry

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We found a novel glycosyltransferase gene having a hypothetical ␤1,4-galactosyltransferase motif (GenBank TM accession number AB081516) by a BLAST search and cloned its full-length open reading frame using the 5-rapid amplification of cDNA ends method. The truncated form was expressed in insect cells as a soluble enzyme. It transferred N-acetylgalactosamine, not galactose, to para-nitrophenyl-␤-glucuronic acid. The N-acetylgalactosamine-glucuronic acid linkage has been identified only in chondroitin sulfate; therefore, we examined its chondroitin elongation and initiation activities. N-Acetylgalactosaminyltransferase activity was observed toward chondroitin poly-and oligosaccharides, chondroitin sulfate oligosaccharides, and linkage tetrasaccharide (GlcA-Gal-Gal-Xyl-O-methoxyphenyl), and the chondroitin polysaccharide and linkage tetrasaccharide were better acceptor substrates than the others. Northern blot analysis and quantitative realtime PCR analysis revealed that its 4-kb transcripts were highly expressed in thyroid and placenta, although they were ubiquitously expressed in various tissues and cells. These results suggest that this enzyme has N-acetylgalactosaminyltransferase activity in both the elongation and initiation of chondroitin sulfate synthesis. Furthermore, we performed enzymatic synthesis of chondroitin pentasaccharide in vitro. In one tube reaction with four enzymes, ␤1,4-galactosyltransferase-VII, ␤1,3-galactosyltransferase-VI, glucuronyltransferase-I, and this enzyme, and a synthetic xylose-peptide acceptor, the structure GalNAc-GlcA-Gal-Gal-Xyl-peptide was constructed. This is the first report of a chondroitin pentasaccharide constructed with recombinant glycosyltransferases in vitro.

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First Isolation of Human UDP-d-Xylose: Proteoglycan Core Protein β-d-Xylosyltransferase Secreted from Cultured JAR Choriocarcinoma Cells

Cited by 70 publications

References 38 publications

Functional Characterization of UDP-apiose Synthases from Bryophytes and Green Algae Provides Insight into the Appearance of Apiose-containing Glycans during Plant Evolution

Functional Characterization of UDP-apiose Synthases from Bryophytes and Green Algae Provides Insight into the Appearance of Apiose-containing Glycans during Plant Evolution

Enzyme interactions in heparan sulfate biosynthesis: Uronosyl 5-epimerase and 2- O -sulfotransferase interact in vivo

Enzymatic Synthesis of Chondroitin with a Novel Chondroitin Sulfate N-Acetylgalactosaminyltransferase That Transfers N-Acetylgalactosamine to Glucuronic Acid in Initiation and Elongation of Chondroitin Sulfate Synthesis

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