Synthesis of a sulfated glycopeptide corresponding to the carbohydrate-protein linkage region of proteoglycans: β-D-GlcA-(1→3){(SO3Na→4)}-β-D-Gal-(1→3)-β-D-Gal-(1→4)-β-D-Xyl-(1→3)-Ser

Goto, Fumitaka; Ogawa, Tomoya

doi:10.1016/s0040-4039(00)61200-5

Cited by 31 publications

(20 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…UDP-[ 3 H]GalNAc (6.3 Ci/mmol) and unlabeled UDPGalNAc were purchased from DuPont NEN and Sigma, respectively. The following linkage tetrasaccharide and hexasaccharide serines were synthesized chemically (11)(12)(13)…”

Section: Methodsmentioning

confidence: 99%

“…Sulfated Gal residues have not been found to date in the linkage region of heparin or heparan sulfate (9,10). In view of these structural variations in the linkage region, we investigated the effects of sulfation in the linkage region on the specificity of GalNAc transferase, which is involved in the biosynthesis of chondroitin sulfate using chemically synthesized linkage tetrasaccharide serines, GlcA␤1-3Gal␤1-3Gal␤1-4Xyl␤1-O-Ser and GlcA␤1-3Gal(4-sulfate)␤1-3Gal ␤1-4Xyl␤1-O-Ser, and hexasaccharide serines, GlcA␤1-3GalNAc␤1-4GlcA␤1-3Gal␤1-3Gal␤1-4Xyl␤1-O-Ser, GlcA␤1-3GalNAc(4-sulfate)␤1-4GlcA␤1-3Gal␤1-3Gal␤1-4Xyl␤1-O-Ser, and GlcA␤1-3GalNAc(4-sulfate)␤1-4GlcA␤1-3Gal(4-sulfate)␤1-3Gal␤1-4Xyl␤1-O-Ser, as acceptor substrates (11)(12)(13). During studies to characterize the enzyme products, we unexpectedly found that the products contained the transferred [ 3 H]GalNAc ␣-linked to the nonreducing terminal GlcA of the acceptor substrates employed.…”

mentioning

confidence: 99%

See 1 more Smart Citation

N-Acetylgalactosamine (GalNAc) Transfer to the Common Carbohydrate-Protein Linkage Region of Sulfated Glycosaminoglycans

Kitagawa

Tanaka

Tsuchida

et al. 1995

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

N-Acetylgalactosamine (GalNAc) Transfer to the Common Carbohydrate-Protein Linkage Region of Sulfated Glycosaminoglycans

Kitagawa

Tanaka

Tsuchida

et al. 1995

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…The purity and concentration of these chemically synthesized substrates were checked by 1 H NMR spectroscopy (15)(16)(17) and high performance liquid chromatography (HPLC) on an amine-bound silica column as described (18), respectively. The following unsaturated linkage tetrasaccharide-and hexasaccharide-serines, and hexasaccharide alditols were isolated from porcine intestinal heparin (9), rat chondrosarcoma chondroitin 4-sulfate (3), whale cartilage chondroitin 4-sulfate (4), and shark cartilage chondroitin 6-sulfate (6, 7) as described:…”

Section: Methodsmentioning

confidence: 99%

“…). The following linkage tri-, tetra-, penta-, and hexasaccharide-serines and a tetrasaccharide-peptide were chemically synthesized (15)(16)(17)…”

mentioning

confidence: 99%

Sulfation of the Galactose Residues in the Glycosaminoglycan-Protein Linkage Region by Recombinant Human Chondroitin 6-O-Sulfotransferase-1

Kitagawa

Tsutsumi

Ikegami-Kuzuhara

et al. 2008

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

6-O-Sulfated galactose residues have been demonstrated in the glycosaminoglycan-protein linkage region GlcUA␤1-3Gal␤1-3Gal␤1-4Xyl␤1-O-Ser isolated from shark cartilage chondroitin 6-sulfate (Sugahara, K., Ohi, Y., Harada, T., de Waard, P., and Vliegenthart, J. F. G. (1992) J. Biol. Chem. 267, 6027-6035). In this study, we investigated whether a recombinant human chondroitin 6-sulfotransferase-1 (C6ST-1) catalyzes the sulfation of C6 on both galactose residues in the linkage region using structurally defined acceptor substrates. The C6ST-1 was expressed as a soluble protein A chimeric form in COS-1 cells and purified using IgG-Sepharose. The purified C6ST-1 utilized the linkage tri-, tetra-, penta-, and hexasaccharide-serines and hexasaccharide alditols, including GlcUA␤1-3GalNAc(4-O-sulfate)␤1-4GlcUA␤1-3Gal(4-Osulfate)␤1-3Gal␤1-4Xyl␤1-O-Ser and ⌬GlcUA␤1-3Gal-NAc(6-O-sulfate)␤1-4GlcUA␤1-3Gal␤1-3Gal(6-O-sulfate)␤1-4Xyl-ol. Identification of the reaction products obtained with the linkage tetra-, penta-, and hexasaccharideserines revealed that the C6ST-1 catalyzed the sulfation of C6 on both galactose residues in the linkage region. Notably, the linkage tetrasaccharide-peptide GlcUA␤1-3Gal␤1-3Gal␤1-4Xyl␤1-O-(Gly)Ser-(Gly-Glu) was a good acceptor substrate for the C6ST-1, suggesting that the sulfation of the galactose residues can occur before the transfer of the first N-acetylhexosamine residue to the linkage tetrasaccharide. In contrast, no incorporation was observed into ⌬GlcUA␤1-3GalNAc(4-O-sulfate)␤1-4GlcUA␤1-3Gal(4-O-sulfate)␤1-3Gal␤1-4Xyl-ol, indicating that an intact xylose is necessary for the transfer of a sulfate to the second sugar residue Gal from the reducing end. These findings clearly demonstrated that the recombinant C6ST-1 catalyzes the sulfation of C6 on both galactose residues in the linkage region in vitro. This is the first identification of the sulfotransferase responsible for the sulfation of galactose residues in the glycosaminoglycanprotein linkage region. Sulfated glycosaminoglycans (GAGs),3 including heparin/ heparan sulfate, chondroitin sulfate, and dermatan sulfate, are covalently bound to Ser residues in the core proteins through the common carbohydrate-protein linkage structure, GlcUA␤1-3Gal␤1-3Gal␤1-4Xyl␤1-O-Ser. Heparin/ heparan sulfate is synthesized once GlcNAc is transferred to the common linkage region, whereas chondroitin sulfate is formed if GalNAc is first added. Therefore, the first hexosamine transfer is critical in determining whether heparin/ heparan sulfate or chondroitin/dermatan sulfate chains are selectively assembled on the common linkage region (for reviews, see Refs. 1 and 2). Although such mechanisms have long been proposed based on data from conventional structural and enzymological studies (2, 3), the molecular mechanisms underlying the selective chain assembly of different GAG chains have not yet been clarified.We have been investigating the structure of the linkage region of various GAGs to search for possible structural differences, which may determine the ch...

show abstract

“…Unsaturated hexasaccharide alditols were isolated from the GAG–protein linkage region of chondroitin sulfate from shark cartilage and bovine nasal septa as reported previously [4,5,21]. Oligosaccharide‐serines were also synthesized chemically [22,23].…”

Section: Methodsmentioning

confidence: 99%

Substrate specificity studies of Flavobacterium chondroitinase C and heparitinases towards the glycosaminoglycan–protein linkage region

Tsuda¹,

Yamada²,

Miyazono³

et al. 1999

European Journal of Biochemistry

Self Cite

View full text Add to dashboard Cite

Bacterial chondroitinases and heparitinases are potentially useful tools for structural studies of chondroitin sulfate and heparin/heparan sulfate. Substrate specificities of Flavobacterium chondroitinase C, as well as heparitinases I and II, towards the glycosaminoglycan±protein linkage region -HexA-HexNAc-GlcA-Gal-Gal-Xyl-Ser (where HexA represents glucuronic acid or iduronic acid and HexNAc represents N-acetylgalactosamine or N-acetylglucosamine) were investigated using various structurally defined oligosaccharides or oligosaccharide-serines derived from the linkage region. In the case of oligosaccharide-serines, they were labeled with a chromophore dimethylaminoazobenzenesulfonyl chloride (DABS-Cl), which stably reacted with the amino group of the serine residue and rendered high absorbance for microanalysis. Chondroitinase C cleaved the GalNAc bond of the pentasaccharides or hexasaccharides derived from the linkage region of chondroitin sulfate chains and tolerated sulfation of the C-4 or C-6 of the GalNAc residue and C-6 of the Gal residues, as well as 2-O-phosphorylation of the Xyl residue. In contrast, it did not act on the GalNAc±GlcA linkage when attached to a 4-O-sulfated Gal residue. Heparitinase I cleaved the innermost glucosaminidic bond of the linkage region oligosaccharide-serines of heparin/ heparan sulfate irrespective of substitution by uronic acid, whereas heparitinase II acted only on the glucosaminidic linkages of the repeating disaccharide region, but not on the innermost glucosaminidic linkage. These defined specificities of chondroitinase C, as well as heparitinases I and II, will be useful for preparation and structural analysis of the linkage oligosaccharides.

show abstract

Synthesis of a sulfated glycopeptide corresponding to the carbohydrate-protein linkage region of proteoglycans: β-D-GlcA-(1→3){(SO3Na→4)}-β-D-Gal-(1→3)-β-D-Gal-(1→4)-β-D-Xyl-(1→3)-Ser

Cited by 31 publications

References 32 publications

N-Acetylgalactosamine (GalNAc) Transfer to the Common Carbohydrate-Protein Linkage Region of Sulfated Glycosaminoglycans

N-Acetylgalactosamine (GalNAc) Transfer to the Common Carbohydrate-Protein Linkage Region of Sulfated Glycosaminoglycans

Sulfation of the Galactose Residues in the Glycosaminoglycan-Protein Linkage Region by Recombinant Human Chondroitin 6-O-Sulfotransferase-1

Substrate specificity studies of Flavobacterium chondroitinase C and heparitinases towards the glycosaminoglycan–protein linkage region

Contact Info

Product

Resources

About