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Structural studies of two populations of keratan sulphate chains from mature bovine articular cartilage

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Abstract

Two discrete peptido-keratan sulphate fragments were isolatedvia chondroitinase ABC and trypsin digestion of a proteoglycan aggregate fraction prepared from bovine femoral head cartilage (six year old animals). The larger fragments (Kav=0.07, CL-6B) contained peptides substituted with several keratan sulphate (KS) chains from the KS-rich region of the proteoglycan and the smaller fragments (Kav=0.5, CL-6B) contained peptides with, perhaps, only one KS chain and the stubs of post-chondroitinase-treated chondroitin sulphate chains.

The two peptido-KS samples and the KS chains derived from these by alkaline borohydride reduction were characterised by13C-NMR spectroscopy. The two populations of KS chains were also examined by chromatography (Sephadex G-75), and keratanase digestion followed by chromatography on Bio-Gel P-10. From the results it was concluded that the KS chains from the two major trypsin-derived peptido-KS fragments had similar sulphation levels, distributions of hydrodynamic sizes and susceptibilities to keratanase.

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Abbreviations

KS:

keratan sulphate

A1:

proteoglycan aggregate

T:

diphenyl carbamyl chloride (DPCC)-trypsintreated

CB:

chondroitinase ABC-treated

C:

chymotrypsin-treated

P:

papain-treated

R:

alkaline borohydride-reduced

TSP:

sodium 3-trimethylsilylpropionate

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Author notes

  1. David J Thornton

    Present address: Department of Biochemistry, University of Manchester, Stopford Building, Oxford Road, M13 9PT, Manchester, U.K.

Authors and Affiliations

  1. Division of Biological Sciences, Institute of Environmental and Biological Sciences, University of Lancaster, Bailrigg, LA1 4YQ, Lancaster, U.K.

    David J Thornton, Haydn G Morris, Gordon H Cockin & Ian A Nieduszynski

  2. Department of Chemistry, University of Lancaster, Bailrigg, LA1 4YA, Lancaster, U.K.

    Thomas N Huckerby

Authors
  1. David J Thornton
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  2. Haydn G Morris
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  3. Gordon H Cockin
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  4. Thomas N Huckerby
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  5. Ian A Nieduszynski
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Thornton, D.J., Morris, H.G., Cockin, G.H. et al. Structural studies of two populations of keratan sulphate chains from mature bovine articular cartilage. Glycoconjugate J 6, 209–218 (1989). https://doi.org/10.1007/BF01050649

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  • DOI: https://doi.org/10.1007/BF01050649

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