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.

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.

scholarly journals Turnover of proteoglycans in articular-cartilage cultures. Characterization of proteoglycans released into the medium

1989 ◽  
Vol 259 (1) ◽  
pp. 21-25 ◽  
Author(s):  
M A Campbell ◽  
C J Handley ◽  
S E D'Souza
Keyword(s):  
Articular Cartilage ◽  
Core Protein ◽  
Chondroitin Sulphate ◽  
Polyacrylamide Gel Electrophoresis ◽  
Keratan Sulphate ◽  
Bovine Articular Cartilage ◽  
Definitive Evidence ◽  
Core Proteins ◽  
The Matrix

By using an e.l.i.s.a. method it was demonstrated that the majority of proteoglycans released into the medium of both control and retinoic acid-treated explant cultures of bovine articular cartilage did not contain a hyaluronate-binding region. This supports our previous findings [Campbell & Handley (1987) Arch. Biochem. Biophys. 258, 143-155] that proteoglycans released into the medium of both cultures were of smaller hydrodynamic size, more polydisperse and unable to form aggregates with hyaluronate. Analysis of 35S-labelled core proteins associated with proteoglycans released into the medium of both cultures by using SDS/polyacrylamide-gel electrophoresis and fluorography indicated the presence of a series of core-protein bands (Mr approx. 300,000, 230,000, 215,000, 200,000, 180,000, 140,000, 135,000, 105,000, 85,000 and 60,000) compared with three core proteins derived from the proteoglycans remaining in the matrix (Mr 300,000, 230,000 and 215,000). Further analysis of the core proteins released into the medium indicated that the larger core proteins associated with medium proteoglycans contain both chondroitin sulphate and keratan sulphate glycosaminoglycans whereas the smaller core proteins contain only chondroitin sulphate chains. These experiments provide definitive evidence that the loss of proteoglycans from the matrix involves proteolytic cleavage at various sites along the proteoglycan core protein.

Oligosaccharides Derived from Bovine Articular Cartilage Keratan Sulfates after Keratanase II Digestion: Implications for Keratan Sulfate Structural Fingerprinting

Biochemistry ◽  
1994 ◽  
Vol 33 (16) ◽  
pp. 4836-4846 ◽  
Author(s):  
Gavin M. Brown ◽  
Thomas N. Huckerby ◽  
Haydn G. Morris ◽  
Beverley L. Abram ◽  
Ian A. Nieduszynski
Keyword(s):  
Articular Cartilage ◽  
Keratan Sulfate ◽  
Bovine Articular Cartilage ◽  
Keratanase Ii

scholarly journals Fucose content of keratan sulphates from bovine articular cartilage

1991 ◽  
Vol 273 (2) ◽  
pp. 307-310 ◽  
Author(s):  
G H Tai ◽  
G M Brown ◽  
H G Morris ◽  
T N Huckerby ◽  
I A Nieduszynski
Keyword(s):  
Molecular Weight ◽  
Articular Cartilage ◽  
Molecular Size ◽  
Gel Permeation Chromatography ◽  
Repeat Sequence ◽  
Keratan Sulphate ◽  
Bovine Articular Cartilage ◽  
Gel Permeation ◽  
Fucose Content ◽  
Galactose Content

Alkaline-borohydride-reduced keratan sulphate chains were isolated from bovine articular cartilage (6-8-year-old animals). Nine keratan sulphate fractions of increasing molecular weight were prepared by gel-permeation chromatography on a calibrated column of TSK 30 XL. The samples were analysed for fucose and galactose contents (% by wt. of keratan sulphate) and fucose/galactose ratio. The fucose content increased with molecular size, but the galactose content remained constant. It was concluded that the alpha(1→3)-linked fucose [Thornton, Morris, Cockin, Huckerby, Nieduszynski, Carlstedt, Hardingham & Ratcliffe (1989) Biochem. J. 260, 277-282] was located within the poly-N-acetyl-lactosamine repeat sequence of articular-cartilage keratan sulphate.

scholarly journals Isolation and characterization of cyanogen bromide peptides from the collagen of bovine articular cartilage

1973 ◽  
Vol 133 (4) ◽  
pp. 615-622 ◽  
Author(s):  
Kalindi Deshmukh ◽  
Marcel E. Nimni
Keyword(s):  
Articular Cartilage ◽  
Total Amino Acid ◽  
Cross Linking ◽  
Amino Acid Residues ◽  
Bovine Articular Cartilage ◽  
Isolation And Characterization ◽  
Peptide Pattern ◽  
Intact Cartilage ◽  
Native Collagen

Significant amounts of native collagen can be extracted from bovine articular cartilage after removal of the acid mucopolysaccharides by controlled proteolysis. The fraction thus solubilized upon denaturation gives rise to three identical α chains. Cleavage of these chains with CNBr generated nine peptides, all of which contain glycine as one-third of their total amino acid residues. Two of the smaller peptides CB-1 and CB-2 contain partially hydroxylated proline. A similar CNBr digest of intact cartilage also gives a series of peptides identical with those obtained from the soluble cartilage collagen. The absence of cross-linking peptides, the fact that only few β components are seen in articular cartilage collagen and the similarity in peptide pattern between the two collagen fractions investigated, suggests that this collagen is stabilized by a different cross-linking mechanism, possibly involving an association with the tissue proteoglycans.

scholarly journals Articular cartilage cultured with interleukin 1. Increased release of link protein, hyaluronate-binding region and other proteoglycan fragments

1986 ◽  
Vol 238 (2) ◽  
pp. 571-580 ◽  
Author(s):  
A Ratcliffe ◽  
J A Tyler ◽  
T E Hardingham
Keyword(s):  
Articular Cartilage ◽  
Interleukin 1 ◽  
Binding Region ◽  
Rapid Loss ◽  
Keratan Sulphate ◽  
Link Protein ◽  
The Matrix ◽  
The Media ◽  
Average Size

Pig articular cartilage was maintained in culture for 3 days with and without porcine interleukin 1. The proteoglycans remaining in the cartilage and those released into the medium were analysed by using radioimmunoassays for the hyaluronate-binding region, link protein and keratan sulphate. In interleukin 1-treated cultures after 3 days there was 38% release of total glycosaminoglycans into the medium, 18% release of binding region, 14% release of link protein and 20% release of keratan sulphate epitope, whereas in control cultures the proportions released were much less (16, 9, 10 and 7% respectively). Characterization of the proteoglycans in the media after 1.5 days and 3 days of culture showed that interleukin 1 promoted the release of proteoglycan of large average size and also the release of link protein and of low-Mr binding region which was unattached to proteoglycan. Both the link protein and binding region released were able to bind to exogenously added hyaluronate, whereas the proteoglycan in the medium was not. The proteoglycans extracted from cultured cartilage were similar to those from fresh cartilage: they contained a high proportion of aggregating proteoglycans and some low-Mr binding region. The proportion of this binding region extracted from the interleukin 1-treated cartilage was increased. The presence of interleukin 1 in the cultures therefore appeared to increase the rate of proteolytic degradation of proteoglycan in the matrix and to lead to a more rapid loss of intact binding region, of link protein and of large proteoglycan fragments into the medium.

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