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Keratanase digestion of keratan sulphates: characterisation of large oligosaccharides from the N-acetyllactosamine repeat sequence and from the non-reducing terminal chain caps.

Huckerby, Thomas N. and Dickenson, John M. and Brown, Gavin M. and Nieduszynski, Ian A. (1995) Keratanase digestion of keratan sulphates: characterisation of large oligosaccharides from the N-acetyllactosamine repeat sequence and from the non-reducing terminal chain caps. Biochimica et Biophysica Acta (BBA) - General Subjects, 1244 (1). pp. 17-29.

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Abstract

Keratan sulfate (KS) chains prepared from both bovine tracheal rings and bovine femoral head cartilage were digested with the enzyme keratanase from Pseudomonas species; large repeat-sequence and non-reducing terminal oligosaccharides were fractionated and purified using high-performance ion-exchange chromatography. The main beta-linked pentasulfated hexasaccharide repeat segment, [R6], GlcNAc(6S)1-1-3Gal(6S)1-4GlcNAc(6S)1-3Gal(6S)1-4GlcNAc(6S)1-3Gal-ol and the asialo beta-linked capping pentasulfated heptasaccharide, [C7], Gal1-4GlcNAc(6S)1-3Gal(6S)1-4GlcNAc(6S)1-3Gal(6S)1-4GlcNAc(6S) 1-3Gal-ol have been completely characterized by high-field NMR spectroscopy using one- and two-dimensional methods. Partial 1H assignments are summarized for three homologous series of higher oligosaccharides: GlcNAc(6S)[1-3Gal(6S)1-4GlcNAc(6S)]2-5(1-3)Gal-0l [R8,R10,R12] Gal1-4GlcNAc(6S)[1-3Gal(6S)1-4GlcNAc(6S)]3-5(1-3)Gal-ol [C9,C11,C13] NeuAc alpha 2-3Gal1-4GlcNAc(6S)[1-3Gal(6S)1-4GlcNAc(6S)]2-4(1-3)Gal-ol [C8,C10,C12] obtained from keratan sulfate by keratanase cleavage. The first shows that the unsulfated galactose residues within the repeat sequence region of KS may be separated by fully sulfated segments which have a wide distribution of lengths. The others, viz. those with sialylated caps, and the related galactose capped asialo-segments (derived from a KS digestion in which the keratanase also exhibited sialidase activity) represent an homologous series of epitopes in which the first internal unsulfated galactose is located at a position which may be up to five or more fully sulfated N-acetyllactosamine disaccharide repeat units along from the non-reducing terminus of the KS polymer.

Item Type: Article
Journal or Publication Title: Biochimica et Biophysica Acta (BBA) - General Subjects
Uncontrolled Keywords: Keratan sulfate oligosaccharide ; NMR ; 1H- ; Repeat sequence ; Chain cap ; Structural assignment ; Keratanase digestion
Subjects: G Geography. Anthropology. Recreation > GE Environmental Sciences
Departments: Faculty of Science and Technology > Lancaster Environment Centre
Faculty of Health and Medicine > Biomedical & Life Sciences
VC's Office
ID Code: 22283
Deposited By: ep_ss_importer
Deposited On: 06 Feb 2009 15:33
Refereed?: Yes
Published?: Published
Last Modified: 26 Jul 2012 15:58
Identification Number:
URI: http://eprints.lancs.ac.uk/id/eprint/22283

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