Constant and variable domains of different disaccharide structure in corneal keratan sulphate chains

Oeben, M; Keller, R.; Stuhlsatz, H. W.; Greiling, H.

doi:10.1042/bj2480085

Cited by 78 publications

(52 citation statements)

References 18 publications

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“…1, panel B) suggests that a critical minimum polymer size is necessary for synthesis of oversulfated regions in the chain interior. Conversely, as suggested by earlier in vitro studies (44,45), keratan sulfate chain elongation may only proceed in conjunction with efficient sulfation, and the undersulfated chains in both types of MCD, and in both tissues examined, were significantly shorter than those isolated from normal tissues. Thus, premature chain termination in the absence of appropriate sulfation may occur if sulfation The migrations of disaccharide glcNAc␤1, 3gal, the trisaccharide gal␤1,4glcNAc ␤1,3gal, and neuA were determined by coelectrophoresis of standards as reported (29) (data not shown).…”

Section: Discussionmentioning

confidence: 55%

Altered Fine Structures of Corneal and Skeletal Keratan Sulfate and Chondroitin/Dermatan Sulfate in Macular Corneal Dystrophy

Plaas

West

Thonar

et al. 2001

Journal of Biological Chemistry

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The content and fine structure of keratan and chondroitin/dermatan sulfate in normal human corneas and corneas affected by macular corneal dystrophies (MCD) types I and II were examined by fluorophore-assisted carbohydrate electrophoresis. Normal tissues (n ‫؍‬ 11) contained 15 g of keratan sulfate and 8 g of chondroitin/dermatan sulfate per mg dry weight. Keratan sulfates consisted of ϳ4% unsulfated, 42% monosulfated, and 54% disulfated disaccharides with number of average chain lengths of ϳ14 disaccharides. Chondroitin/ dermatan sulfates were significantly longer, ϳ40 disaccharides per chain, and consisted of ϳ64% unsulfated, 28% 4-sulfated, and 8% 6-sulfated disaccharides. The fine structural parameters were altered in all diseased tissues. Keratan sulfate chain size was reduced to 3-4 disaccharides; chain sulfation was absent in MCD type I corneas and cartilages, and sulfation of both GlcNAc and Gal was significantly reduced in MCD type II. Chondroitin/dermatan sulfate chain sizes were also decreased in all diseased corneas to ϳ15 disaccharides, and the contents of 4-and 6-sulfated disaccharides were proportionally increased. Tissue concentrations (nanomole of chains per mg dry weight) of all glycosaminoglycan types were affected in the disease types. Keratan sulfate chain concentrations were reduced by ϳ24 and ϳ75% in type I corneas and cartilages, respectively, and by ϳ50% in type II corneas. Conversely, chondroitin/ dermatan sulfate chain concentrations were increased by 60 -70% in types I and II corneas. Such changes imply a modified tissue content of individual proteoglycans and/or an altered efficiency of chain substitution on the core proteins. Together with the finding that hyaluronan, not normally present in healthy adult corneas, was also detected in both disease subtypes, the data support the conclusion that a wide range of keratocyte-specific proteoglycan and glycosaminoglycan remodeling processes are activated during degeneration of the stromal matrix in the macular corneal dystrophies.

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Section: Discussionmentioning

confidence: 55%

Altered Fine Structures of Corneal and Skeletal Keratan Sulfate and Chondroitin/Dermatan Sulfate in Macular Corneal Dystrophy

Plaas

West

Thonar

et al. 2001

Journal of Biological Chemistry

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“…Also, although information about the levels of galactose sulphation at the chain cap is available none is available about the levels along the repeat region. However, previous work has reported decreasing levels of galactose sulphation along the repeat region towards the reducing terminus (the protein core) (Lauder et al, 1994;Oeben et a]., 1987).…”

Section: Discussionmentioning

confidence: 87%

“…The 0-linked keratan sulphate chains from bovine nasal septum are also short, having a repeat region of about seven to nine disaccharides (Stuhlstatz et al, 1989). However, Oeben et al (1987) found Nlinked keratan sulphate chains from pig cornea with repeat regions of 10-52 disaccharides. These workers found no N-acetylneuraminic acid chain caps so the chains may be even longer in vivo.…”

Section: Discussionmentioning

confidence: 97%

The Structure of the Keratan Sulphate Chains Attached to Fibromodulin Isolated from Articular Cartilage

Lauder

Huckerby

Nieduszynski

1996

European Journal of Biochemistry

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Fibromodulin has been isolated from bovine and equine articular cartilage and the attached keratan sulphate chains subjected to digestion by keratanase 11. The oligosaccharides generated have been reduced and subsequently isolated by strong anion-exchange chromatography. Their structures have been determined by high-field 'H-NMR spectroscopy and high-pH anion-exchange chromatography.Both a(2-6)-and a(2-3)-linked N-acetylneuraminic acid have been found in the capping oligosaccharides, and, fucose which is a(l-3)-linked to N-acetylglucosamine has been found as a branch in both repeat region and capping oligosaccharides. These data demonstrate that there are fundamental differences between the structures present in the N-linked keratan sulphate chains attached to fibromodulin from articular cartilage and those from tracheal cartilage, which lack both a(2-6)-linked N-acetylneuraminic acid and a( 1 -3)-linked fucose.It has been confirmed that the keratan sulphate chains are short, being only eight or nine disaccharides in length. Very significant differences in the levels of galactose sulphation have been identified at the non-reducing end of the chain. The galactose residue adjacent to the non-reducing cap is sulphated in only 1-3% of chains, compared with a sulphation level of over 40% closer to the reducing end. This highlights the difference between the chain termini and the repeat region in terms of structure and points to the potential for functional importance.The repeat region and capping fragments of the N-linked keratan sulphates from bovine and equine articular cartilage fibromodulin have been found to have the following general structure : NeuAc-(a2-3/6)Ga1[6SO;] (PI-4) GlcNAc6SO; (pl-3) Gal [6SO,] (Pl-4) { [Fuc(al-3)],,GlcNAc6SO~ (pl-3)Gal-

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“…This is identical structure to that we found. Oeben et al (40), however, reported a biantennary complex type structure that has a sialylgalactosyl N-acetyllactosamine and e), pcDNA3-hCGn6ST (lanes b and f), pcDNA3-hKSG6ST (lanes c and g), and both pcDNA3-hCGn6ST and pcDNA3-hKSG6ST vectors (lanes d and h). Membrane protein fractions were prepared from those transfectants and separated by SDS-PAGE followed by Coomassie Blue staining (lanes a-d) or Western blotting using an anti-keratan sulfate antibody, 5D4 (lanes e-h).…”

Section: Discussionmentioning

confidence: 97%

Human Corneal GlcNAc 6-O-Sulfotransferase and Mouse Intestinal GlcNAc 6-O-Sulfotransferase Both Produce Keratan Sulfate

Akama

Nakayama

Nishida

et al. 2001

Journal of Biological Chemistry

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Human corneal N-acetylglucosamine 6-O-sulfotransferase (hCGn6ST) has been identified by the positional candidate approach as the gene responsible for macular corneal dystrophy (MCD). Because of its high homology to carbohydrate sulfotransferases and the presence of mutations of this gene in MCD patients who lack sulfated keratan sulfate in the cornea and serum, hCGn6ST protein is thought to be a sulfotransferase that catalyzes sulfation of GlcNAc in keratan sulfate. In this report, we analyzed the enzymatic activity of hCGn6ST by expressing it in cultured cells. A lysate prepared from HeLa cells transfected with an intact form of hCGn6ST cDNA or culture medium from cells transfected with a secreted form of hCGn6ST cDNA showed an activity of transferring sulfate to C-6 of GlcNAc of synthetic oligosaccharide substrates in vitro. When hCGn6ST was expressed together with human keratan sulfate Gal-6-sulfotransferase (hKSG6ST), HeLa cells produced highly sulfated carbohydrate detected by an anti-keratan sulfate antibody 5D4. These results indicate that hCGn6ST transfers sulfate to C-6 of GlcNAc in keratan sulfate. Amino acid substitutions in hCGn6ST identical to changes resulting from missense mutations found in MCD patients abolished enzymatic activity. Moreover, mouse intestinal GlcNAc 6-O-sulfotransferase had the same activity as hCGn6ST. This observation suggests that mouse intestinal GlcNAc 6-O-sulfotransferase is the orthologue of hCGn6ST and functions as a sulfotransferase to produce keratan sulfate in the cornea.

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Constant and variable domains of different disaccharide structure in corneal keratan sulphate chains

Cited by 78 publications

References 18 publications

Altered Fine Structures of Corneal and Skeletal Keratan Sulfate and Chondroitin/Dermatan Sulfate in Macular Corneal Dystrophy

Altered Fine Structures of Corneal and Skeletal Keratan Sulfate and Chondroitin/Dermatan Sulfate in Macular Corneal Dystrophy

The Structure of the Keratan Sulphate Chains Attached to Fibromodulin Isolated from Articular Cartilage

Human Corneal GlcNAc 6-O-Sulfotransferase and Mouse Intestinal GlcNAc 6-O-Sulfotransferase Both Produce Keratan Sulfate

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