A sub-population of keratan sulphates derived from bovine articular cartilage is capped with alpha(2-6)-linked N-acetylneuraminic acid residues. Affinity chromatography using immobilized Sambucus nigra lectin and characterization using 1H n.m.r. spectroscopy.
Alkaline borohydride-reduced keratan sulphate (KS) chains derived from bovine femoral head cartilage were fractionated by lectin affinity chromatography with Sambucus nigra agglutinin (SNA) into binding and non-binding populations. Analysis of the SNA-binding and non-binding KS chains using 600 MHz 1H n.m.r. spectroscopy showed that the former population contained alpha(2-6)-N-acetylneuraminic acid residues and the latter contained primarily alpha(2-3)-N-acetylneuraminic acid residues as chain terminators. Both populations contained a similar proportion of alpha(2-3)-N-acetylneuraminic acid residues within their protein-linkage regions, and similar sulphation and fucosylation levels. Analysis of these two fractions by gel-permeation chromatography (g.p.c.) on a TSK-30 XL column showed them to have the same size distributions. It was concluded from the n.m.r. spectra and g.p.c. data that the populations differed primarily in the mode of linkage of the chain-terminating sialic acids.
Tai
G H
GH
Division of Biological Sciences, Institute of Environmental and Biological Sciences, Lancaster University, Bailrigg, U.K.
Morris
H G
HG
Brown
G M
GM
Huckerby
T N
TN
Nieduszynski
I A
IA
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002358:Cartilage, Articular / Q000737:chemistry*; D002417:Cattle; D002846:Chromatography, Affinity; D007632:Keratan Sulfate / Q000737:chemistry* / Q000302:isolation & purification; D037102:Lectins; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid; D037121:Plant Lectins; D054788:Ribosome Inactivating Proteins; D012794:Sialic Acids / Q000032:analysis*
D037102:Lectins; D037121:Plant Lectins; C050600:Sambucus nigra lectins; D012794:Sialic Acids; D007632:Keratan Sulfate; D054788:Ribosome Inactivating Proteins; D019158:N-Acetylneuraminic Acid