Peptido-keratan sulphate fragments were isolated from the nucleus pulposus of bovine intervertebral discs (2-year-old animals) after digestion with chondroitin ABC lyase followed by digestion with diphenylcarbamoyl chloride-treated trypsin of A1D1 proteoglycans and gel-permeation chromatography on Sepharose CL-6B. The peptido-keratan sulphate fragments were subjected to alkaline borohydride reduction. The reduced chains were treated with keratanase in the presence of the sialidase inhibitor 2,3-dehydro-2-deoxy-N-acetylneuraminic acid, and the digest was subjected to alkaline borohydride reduction. This produced oligosaccharides with galactitol at their reducing ends. This reduced digest was chromatographed on a Nucleosil 5 SB anion-exchange column and individual oligosaccharides were isolated. One of these was shown by 600 MHz 1H-n.m.r. spectroscopy to have the following structure: NeuAc alpha 2-6Gal beta 1-4GlcNAc(6-SO4)beta 1-3Gal-ol The structure of this oligosaccharide shows that keratan sulphate chains from bovine intervertebral disc have non-reducing termini with N-acetylneuraminic acid linked alpha(2----6) as well as alpha(2----3) to an unsulphated galactose.
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February 1992
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Research Article|
February 15 1992
Skeletal keratan sulphate chains isolated from bovine intervertebral disc may terminate in α(2----6)-linked N-acetylneuraminic acid
J M Dickenson;
J M Dickenson
*Division of Biological Sciences, Institute of Environmental and Biological Sciences, Lancaster University, Bailrigg, Lancaster LA1 4YQ
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T N Huckerby;
T N Huckerby
†The Polymer Centre, Lancaster University, Bailrigg, Lancaster LA1 4YA, U.K.
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I A Nieduszynski
I A Nieduszynski
*Division of Biological Sciences, Institute of Environmental and Biological Sciences, Lancaster University, Bailrigg, Lancaster LA1 4YQ
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Publisher: Portland Press Ltd
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 1992 The Biochemical Society, London
1992
Biochem J (1992) 282 (1): 267–271.
Citation
J M Dickenson, T N Huckerby, I A Nieduszynski; Skeletal keratan sulphate chains isolated from bovine intervertebral disc may terminate in α(2----6)-linked N-acetylneuraminic acid. Biochem J 15 February 1992; 282 (1): 267–271. doi: https://doi.org/10.1042/bj2820267
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