scholarly journals Age-related changes in the structure of the keratan sulphate chains attached to fibromodulin isolated from articular cartilage

1998 ◽  
Vol 330 (2) ◽  
pp. 753-757 ◽  
Author(s):  
M. Robert LAUDER ◽  
N. Thomas HUCKERBY ◽  
A. Ian NIEDUSZYNSKI ◽  
H. K. Anna PLAAS
Keyword(s):  
Articular Cartilage ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Keratan Sulphate ◽  
Bovine Articular Cartilage ◽  
Acetylneuraminic Acid ◽  
Age Related ◽  
Keratanase Ii ◽  
Age Related Changes ◽  
Related Increase

Bovine articular cartilage fibromodulin has been isolated from animals aged 3 months to 8 years, and the attached keratan sulphate (KS) chains digested with keratanase II. The oligosaccharides generated have been reduced, examined by high-pH anion-exchange chromatography and their structures identified by comparison with standards. It has been shown that in fibromodulin from young articular cartilage, the KS chains do not possess either non-reducing terminal (α2-6)-linked N-acetylneuraminic acid or fucose (α1-3)-linked to sulphated N-acetylglucosamine residues. However, an age-related increase has been observed in the abundance of both (α2-6)-linked N-acetylneuraminic acid and (α1-3)-linked fucose, neither of which is found in KS isolated from non-articular cartilage, irrespective of the age of the source. Interestingly, the KS chain length remains constant as a function of age, which possibly relates to a role in collagen fibril assembly. In addition, no significant age-related changes were identified in levels of galactose sulphation.

Age-related changes in the sulphation of the chondroitin sulphate linkage region from human articular cartilage aggrecan

2001 ◽  
Vol 358 (2) ◽  
pp. 523-528 ◽  
Author(s):  
Robert M. LAUDER ◽  
Thomas N. HUCKERBY ◽  
Gavin M. BROWN ◽  
Michael T. BAYLISS ◽  
Ian A. NIEDUSZYNSKI
Keyword(s):  
Articular Cartilage ◽  
Chondroitin Sulphate ◽  
Basic Structure ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Size Exclusion ◽  
Human Articular Cartilage ◽  
Linkage Region ◽  
Age Related ◽  
Age Related Changes

The chondroitin sulphate (CS) linkage regions have been isolated from human articular cartilage aggrecan (from 10- to 72-year-olds) by chondroitin ABC endolyase digestion and size-exclusion chromatography. Linkage region hexasaccharides have been characterized and their abundance estimated by high-pH anion-exchange chromatography. The basic structure for the CS linkage region oligosaccharides identified from human aggrecan is as follows: ΔUA(β1–3)GalNAc[0S/4S/6S](β1–4)GlcA(β1–3)Gal[0S/6S](β1–3)Gal(β1–4)Xyl, where ΔUA represents 4,5-unsaturated hexuronic acid, 4S and 6S represent an O-ester sulphate group on C-4 and C-6 respectively, and 0S represents zero sulphation. There are significant age-related changes in the abundance of the various N-acetylgalactosamine (GalNAc) sulphation forms identified, occurring up to approx. 20 years old. During the period from 10 to 20 years old the level of GalNAc 6-sulphation at the linkage region increases from approx. 43% to approx. 75%, while there is a corresponding reduction in unsulphated (approx. 30% to approx. 20%) and 4-sulphated (approx. 25% to approx. 6%) GalNAc residues. There is also an increase in the incidence of linkage region galactose 6-sulphation (approx. 2% to approx. 10%) which was only observed in linkage regions with GalNAc 6-sulphation. Beyond 20 years old there are few changes in the relative abundance of these GalNAc sulphation variants; however, there is a slight increase in the abundance of 6-sulphation between approx. 20 years old and approx. 40 years old and a slight decrease in its abundance beyond approx. 40 years old. Our data show that in the majority of chains from tissues of all ages the GalNAc residue closest to the linkage region is 6-sulphated, but the level of GalNAc 6-sulphation within the linkage region is lower than the average level observed within the repeat region.

Age-Related Changes in the Synthesis of Matrix Macromolecules by Bovine Articular Cartilage

1989 ◽  
Vol 19 (2-4) ◽  
pp. 121-133 ◽  
Author(s):  
Philippe Front ◽  
France Aprile ◽  
Dragoslav R. Mitrovic ◽  
David A. Swann
Keyword(s):  
Articular Cartilage ◽  
Bovine Articular Cartilage ◽  
Age Related ◽  
Age Related Changes

scholarly journals Structural and immunological studies of keratan sulphates from mature bovine articular cartilage

1989 ◽  
Vol 260 (1) ◽  
pp. 277-282 ◽  
Author(s):  
D J Thornton ◽  
H G Morris ◽  
G H Cockin ◽  
T N Huckerby ◽  
I A Nieduszynski ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Articular Cartilage ◽  
High Performance ◽  
Gel Permeation Chromatography ◽  
Spectroscopic Studies ◽  
Progressive Increase ◽  
Exchange Chromatography ◽  
Keratan Sulphate ◽  
Bovine Articular Cartilage ◽  
Femoral Head Cartilage

Two populations of alkaline-borohydride-reduced keratan sulphate (KS) chains were prepared from the two peptido-keratan sulphate trypsin fragments of proteoglycan aggregates isolated from bovine femoral head cartilage (6-year-old animals). Each population was separated by high-performance ion-exchange chromatography on a Pharmacia Mono-Q column into eight pools, Q1-Q8. These were analysed by gel permeation chromatography, radioimmunoassay with the monoclonal antibody MZ15, and 500 MHz 1H n.m.r. spectroscopy. Upon chromatography on Sephadex G-75 the Mono-Q fractions were shown to increase in hydrodynamic size progressively from Q1 to Q8 for both KS populations. For each population the strongest antigenic response with the anti-KS monoclonal antibody MZ15 was expressed by the two fractions of greatest size and charge density, Q7 and Q8. Proton n.m.r. spectroscopic studies on the two series of fractions demonstrated: (i) a progressive increase in the level of galactose sulphation from Q1 to Q8, (ii) the presence of approximately one alpha(1-3)-linked fucose residue per chain in every sample, and (iii) the presence of N-acetylneuraminic acids in three discrete environments, two alpha(2-3)- and one alpha(2-6)-linked in every sample. The results are discussed in terms of a possible heterogeneity in the carbohydrate-protein linkage region of keratan sulphates from bovine articular cartilage.

scholarly journals Structure of the keratan sulphate chains attached to fibromodulin isolated from bovine tracheal cartilage. Oligosaccharides generated by keratanase digestion

1994 ◽  
Vol 302 (2) ◽  
pp. 417-423 ◽  
Author(s):  
R M Lauder ◽  
T N Huckerby ◽  
I A Nieduszynski
Keyword(s):  
High Field ◽  
General Structure ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Repeat Region ◽  
Keratan Sulphate ◽  
Tracheal Cartilage ◽  
Acetylneuraminic Acid ◽  
Strong Anion Exchange ◽  
Structure Formula

The structure of the repeat region and chain caps of the N-linked keratan sulphate chains attached to bovine tracheal cartilage fibromodulin has been examined. The chains were fragmented by keratanase digestion, the resultant oligosaccharides isolated by strong anion-exchange chromatography, and their structures determined using high-field 1H-n.m.r. spectroscopy. The chains were found to possess the following general structure: [formula: see text] All of the capping oligosaccharides isolated terminate with alpha(2-3)-linked N-acetylneuraminic acid. No alpha(2-6)-linked N-acetylneuraminic acid chain terminators, nor any fucose, alpha (1-3)-linked to N-acetylglucosamine along the repeat region, were detected. This work demonstrates that the structure of the repeat region and chain caps of N-linked keratan sulphate attached to fibromodulin isolated from bovine tracheal cartilage is identical with that of O-linked keratan sulphate chains attached to aggrecan derived from non-articular cartilage.

scholarly journals Characterization of a non-reducing terminal fragment from bovine articular cartilage keratan sulphates containing α(2-3)-linked sialic acid and α(1-3)-linked fucose. A sulphated variant of the VIM-2 epitope

1996 ◽  
Vol 319 (1) ◽  
pp. 137-141 ◽  
Author(s):  
Gavin M. BROWN ◽  
Thomas N. HUCKERBY ◽  
Beverley L. ABRAM ◽  
Ian A. NIEDUSZYNSKI
Keyword(s):  
Articular Cartilage ◽  
Sialic Acid ◽  
Spectroscopic Analysis ◽  
Anion Exchange Column ◽  
Keratan Sulphate ◽  
Bovine Articular Cartilage ◽  
Sialyl Lex ◽  
Strong Anion Exchange ◽  
Keratanase Ii

Alkaline-borohydride-reduced keratan sulphate chains were isolated from bovine articular cartilage (6–8-year-old animals) and digested with keratanase II, an endo-β-N-acetylglucosaminidase. The resulting oligosaccharides were borohydride-reduced and fractionated on a strong anion-exchange column. 1H-NMR spectroscopic analysis of the products revealed one containing both α(2-3)-linked sialic acid and α(1-3)-linked fucose which was shown to have the structure (I) shown. This structure is a sulphated variant of the VIM-2 epitope (CD65), a putative ligand of E-selectin. No oligosaccharide containing the sialyl-Lex structure [NeuAcα2-3Galβ1-4(Fucα1-3)GlcNAcβ1-] was identified in this study.

Skeletal keratan sulphate structural analysis using keratanase II digestion followed by high-performance anion-exchange chromatography

Glycobiology ◽  
1995 ◽  
Vol 5 (3) ◽  
pp. 311-317 ◽  
Author(s):  
Gavin M. Brown ◽  
Ian A. Nieduszynski ◽  
Haydn G. Morris ◽  
Beverley L. Abram ◽  
Thomas N. Huckerby ◽  
...  
Keyword(s):  
Structural Analysis ◽  
Anion Exchange ◽  
High Performance ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Keratan Sulphate ◽  
Keratanase Ii

Increased incidence of unsulphated and 4-sulphated residues in the chondroitin sulphate linkage region observed by high-pH anion-exchange chromatography

2000 ◽  
Vol 347 (2) ◽  
pp. 339-348 ◽  
Author(s):  
Robert M. LAUDER ◽  
Thomas N. HUCKERBY ◽  
Ian A. NIEDUSZYNSKI
Keyword(s):  
Articular Cartilage ◽  
Anion Exchange ◽  
Chondroitin Sulphate ◽  
Basic Structure ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
High Ph ◽  
Tracheal Cartilage ◽  
Bovine Articular Cartilage ◽  
Linkage Region

We report the isolation, characterization and quantification of five octasaccharides, four hexasaccharides and two tetrasaccharides, derived from the chondroitin sulphate (CS) linkage region of 6-8-year-old bovine articular cartilage aggrecan, following digestion with chondroitin ABC endolyase. Using a novel high-pH anion-exchange chromatography (HPAEC) method, in conjunction with one- and two-dimensional 1H-NMR spectroscopy, we have identified the following basic structure for the CS linkage region of aggrecan: ∆UA(β1-3)GalNAc[0S/4S/6S](β1-4)GlcA(β1-3)GalNAc[0S/4S/6S](β1-4)GlcA(β1-3)Gal[0S/6S](β1-3)Gal(β1-4)Xyl, where ∆UA represents 4,5-unsaturated hexuronic acid, and 4S and 6S represent an O-ester sulphate group on C-4 and C-6 respectively. The octa-, hexa- and tetra-saccharide linkage region fragments were used to develop a HPAEC fingerprinting method, with detection at A232 nm, and a linear response to approx. 0.1 nmol of substance. The sulphation patterns of CS linkage regions, of up to octasaccharide in size, from articular and tracheal cartilage aggrecan were examined. The results show that in articular cartilage, for the majority (53%) of octasaccharides the 2-deoxy-2-N-acetyl amino-D-galactose (GalNAc) residues closest to the linkage region are both 6-sulphated; however, in a significant portion (34%), one or more of these GalNAc residues are unsulphated, and in 8% both are unsulphated. Approximately 10-18% of the chains have a 4-sulphated GalNAc in the first disaccharide, and 12% have a sulphated linkage region Gal residue. No evidence was found for uronic acid sulphation. These data show that there is a significant increase in the incidence of unsulphated and 4-sulphated GalNAc residues adjacent to the linkage region compared with the rest of the chain. Bovine tracheal cartilage linkage regions displayed very similar sulphation profiles to those from articular cartilage, despite the presence of a higher level of GalNAc 4-sulphation within the repeat region of the main CS chain.

scholarly journals Degradation of articular cartilage keratan sulphates using hydrazinolysis and nitrous acid. Environment of fucose residues

1992 ◽  
Vol 286 (1) ◽  
pp. 235-241 ◽  
Author(s):  
G M Brown ◽  
T N Huckerby ◽  
H G Morris ◽  
I A Nieduszynski
Keyword(s):  
Articular Cartilage ◽  
Nitrous Acid ◽  
High Field ◽  
Ion Exchange Chromatography ◽  
Repeat Sequence ◽  
Exchange Chromatography ◽  
Keratan Sulphate ◽  
Bovine Articular Cartilage ◽  
Acid Environment ◽  
Fucose Residue

Alkaline borohydride-reduced keratan sulphate (KS) chains from bovine articular cartilage (6-8-year-old animals) were fragmented by an anhydrous hydrazine/nitrous acid procedure, previously used on KS by Hopwood & Elliott to isolate the major disaccharides from the poly-N-acetyl-lactosamine repeat sequence [Hopwood & Elliott (1983) Carbohydr. Res. 117, 263-274]. The resulting oligosaccharides were reduced with NaB3H4 or NaBH4 and subjected to ion-exchange chromatography on a Nucleosil 5SB column. In addition to the major disaccharides, two fucose-containing oligosaccharides were examined by high-field 1H n.m.r. spectroscopy, and shown to have the following structures (where AnManOH is 2,5-anhydro-D-mannitol): [formula: see text] It is evident that the presence of fucose protects the N-acetylglucosamine residue from de-N-acetylation, and therefore fragments are produced which preserve the immediate environment of the fucose residue. It may be of biosynthetic significance that these two oligosaccharides contain an unsulphated galactose on the non-reducing side of the fucose residue. The hydrazine/nitrous acid/NaB3H4 method followed by h.p.l.c. provides a sensitive fingerprinting technique for the assay of KS composition and sub-populations.

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