title: Structural and immunological studies of keratan sulphates from mature bovine articular cartilage.
abstract: 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.
authors.affiliation: Department of Biological Sciences, University of Lancaster, U.K.
authors.forename: D J
authors.initials: DJ
authors.lastname: Thornton
authors.forename: H G
authors.initials: HG
authors.lastname: Morris
authors.forename: G H
authors.initials: GH
authors.lastname: Cockin
authors.forename: T N
authors.initials: TN
authors.lastname: Huckerby
authors.forename: I A
authors.initials: IA
authors.lastname: Nieduszynski
authors.forename: I
authors.initials: I
authors.lastname: Carlstedt
authors.forename: T E
authors.initials: TE
authors.lastname: Hardingham
authors.forename: A
authors.initials: A
authors.lastname: Ratcliffe
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002358:Cartilage, Articular / Q000032:analysis*; D002417:Cattle; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006025:Glycosaminoglycans / Q000276:immunology*; D007632:Keratan Sulfate / Q000276:immunology* / Q000302:isolation & purification; D009682:Magnetic Resonance Spectroscopy
chemical_list: D000911:Antibodies, Monoclonal; D006025:Glycosaminoglycans; D007632:Keratan Sulfate