The repeat region and chain caps of the N-linked keratan sulphates attached to bovine tracheal cartilage fibromodulin were fragmented by digestion with keratanase II, and the oligosaccharides generated were isolated by strong anion-exchange chromatography. Each of these oligosaccharides has been examined by both HPAE chromatography and high field H-1-NMR spectroscopy.

All of the capping oligosaccharides isolated terminated with alpha(2-3)-linked N-acetyl-neuraminic acid, and neither alpha(2-6)-linked N-acetyl-neuraminic acid chain terminators, nor fucose alpha(1-3)-linked to N-acetylglucosamine were found. The keratan sulphate chains were short, with average lengths of five to seven disaccharides, and the level of galactose sulphation varied along the length of the chain.

The repeat region and chain cap were confirmed as having the following general structure:

[GRAPHICS]

This study has identified a novel structure in fibromodulin, namely a cap containing a sulphated galactose adjacent to a non-reducing terminal N-acetyl-neuraminic acid. We have also confirmed that the general structure of the repeat units and chain caps of N-linked keratan sulphate attached to fibromodulin isolated from bovine tracheal cartilage, is similar to that of O-linked keratan sulphate chains attached to aggrecan from non-articular cartilage. However, there are important differences in chain lengths and sulphation patterns.