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

Biochimica et Biophysica Acta (BBA) - General Subjects
Publication Date
DOI: 10.1016/0304-4165(94)00180-6
  • Keratan Sulfate Oligosaccharide
  • Nmr
  • 1H-
  • Repeat Sequence
  • Chain Cap
  • Structural Assignment
  • Keratanase Digestion
  • Biology


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, β-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 β-linked capping pentasulfated heptasaccharide, [ C7], Gal 1–4GlcNAc(6S) 1–3Ga1(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)[ 1–3 Gal(6S) 1–4GlcNAc(6S)] 3–5 1–3Gal-ol [ R8,R10,R12] Gal 1–4GlcNAc(6S)[ 1–3Gal(6S) 1–4GlcNAc(6S)] 3–5 1–3Gal-ol [ C9,C11,C13] NeuAcα 2–3 Gal 1–4 G1cNAc(6 S)[ 1–3 Gal(6 S) 1–4 GlcNAc(6 S)] 2–4 1–3Gal-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.

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