Oligosaccharides derived by keratanase II digestion of bovine articular cartilage keratan sulphates

Eur J Biochem. 1994 Sep 1;224(2):281-308. doi: 10.1111/j.1432-1033.1994.00281.x.

Abstract

Alkaline borohydride-reduced keratan sulphate chains from bovine articular cartilage (6-8-year-old animals) were subjected to a limit digest with the enzyme keratanase II. Using 1H-NMR spectroscopy, 25 reduced oligosaccharides deriving from keratan sulphate were shown to have the following structures [GlcNAc(6S)-ol represents N-acetylglucosaminitol 6-O-sulphate]: Gal beta 1-4-GlcNAc(6S)-ol, Gal beta 1-4GlcNAc(6S)beta 1-3Gal beta 1-4GlcNAc(6S)-ol, Gal(6S)beta 1-4GlcNAc(6S)-ol, Gal-(6S)beta 1-4GlcNAc(6S) beta 1-3Gal beta 1-4GlcNAc(6S)-ol, Gal beta 1-4GlcNAc(6S)beta 1-3Gal(6S)beta 1-4GlcNAc(6S)-ol, Gal(6S)beta 1-4GlcNAc(6S)beta 1-3Gal(6S)1-4GlcNAc(6S)-ol, Gal beta 1-4(Fuc alpha 1-3)GlcNAc(6S)-ol, Gal beta 1-4-(Fuc alpha 1-3)GlcNAc(6S)beta1-3Gal beta 1-4(Fuc alpha 1-3)GlcNAc(6S)-ol, Gal beta 1-4GlcNAc(6S)beta 1-3Gal beta 1-4(Fuc alpha 1-3)-GlcNAc(6S)-ol, Gal beta 1-4(Fuc alpha 1-3)GlcNAc(6S)beta 1-3Gal beta 1-4GlcNAc(6S)-ol, Gal(6S) beta 1-4GlcNAc-(6S)beta 1-3Gal beta 1-4(Fuc alpha 1-3)GlcNAc(6S)-ol, Gal beta 1-4(Fuc alpha 1-3)GlcNAc(6S)beta 1-3Gal(6S)1-4GlcNAc(6S)-ol, Gal beta 1-4GlcNAc(6S)beta 1-6(Gal beta 1-3)GalNAc-ol, Gal beta 1-4GlcNAc(6S) beta1-6(NeuAc2-3Gal beta 1-3)Gal-NAc-ol, Gal beta 1-4GlcNAc(6S)beta 1-3Gal beta 1-4GlcNAc(6S)beta 1-6(Gal beta 1-3) GalNAc-ol, Gal(6S)beta 1-4GlcNAc-(6S)beta 1-6(Gal beta 1-3)GalNAc-ol, Gal beta 1-4GlcNAc(6S)beta 1-3Gal beta 1-4GlcNAc(6S)beta 1-6(NeuAc2-3Gal beta 1-3)-GalNAc-ol, Gal(6S)beta 1-4GlcNAc(6S)beta 1-6(NeuAc alpha 2-3Gal beta 1-3)GalNAc-ol, Gal(6S) beta 1-4GlcNAc-(6S)beta 1-3Gal beta 1-4GlcNAc(6S)beta 1-6(Gal beta 1-3)GalNAc- ol, Gal(6S)beta 1-4GlcNAc(6S)beta 1-3Gal beta 1-4GlcNAc-(6S)beta 1-6(NeuAc alpha 2-3Gal beta 1-3)GalNAc-ol, NeuAc alpha 2-6Gal beta 1-4GlcNAc(6S)beta 1-3Gal beta 1-4GlcNAc(6S)-ol, NeuAc alpha 2-3Gal beta 1-4GlcNAc(6S)beta 1-3Gal beta 1-4GlcNAc(6S)-ol, NeuAc alpha 2-6Gal beta 1-4GlcNAc(6S)beta 1-3Gal-(6S)beta 1-4GlcNAc(6S)-ol, NeuAc alpha 2-3Gal beta 1-4GlcNAc(6S)beta 1-3Gal(6S)beta 1-4GlcNAc(6S)-ol and Neu-Ac alpha 2-3Gal(6S)beta 1-4GlcNAc(6S)beta 1-3Gal(6S beta)1-4GlcNAc(6S)-ol. Proton chemical shifts for these oligosaccharides were assigned using one- and two-dimensional NMR spectroscopic methods. These results confirm the findings of Nakazawa et al. [Nakazawa, K., Ito, M., Yamagata, T. and Suzuki, S. (1989) in Keratan sulphate: chemistry, biology and chemical pathology (Greiling, H. and Scott, J.E., eds) pp. 99-110, The Biochemical Society, London], namely that keratanase II cleaves the O-glycosidic bond of a beta(1-3)-linked 6-O-sulphated N-acetylglucosamine.(ABSTRACT TRUNCATED AT 400 WORDS)

MeSH terms

  • Acetylglucosaminidase / metabolism*
  • Animals
  • Bacillus / enzymology
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Cartilage, Articular / chemistry*
  • Cattle
  • Chromatography, Ion Exchange
  • Hydrolysis
  • Keratan Sulfate / chemistry*
  • Keratan Sulfate / isolation & purification
  • Magnetic Resonance Spectroscopy
  • Molecular Sequence Data
  • Monosaccharides / analysis
  • Oligosaccharides / chemistry*
  • Oligosaccharides / isolation & purification
  • Oligosaccharides / metabolism
  • Sialic Acids / analysis

Substances

  • Monosaccharides
  • Oligosaccharides
  • Sialic Acids
  • Keratan Sulfate
  • keratanase II
  • Acetylglucosaminidase