Structural and immunological studies of keratan sulphates from mature bovine articular 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.
Thornton
D J
DJ
Department of Biological Sciences, University of Lancaster, U.K.
Morris
H G
HG
Cockin
G H
GH
Huckerby
T N
TN
Nieduszynski
I A
IA
Carlstedt
I
I
Hardingham
T E
TE
Ratcliffe
A
A
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
D000911:Antibodies, Monoclonal; D006025:Glycosaminoglycans; D007632:Keratan Sulfate