Oligosaccharides derived by keratanase II digestion of bovine articular cartilage keratan sulphates.
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)
Brown
G M
GM
Division of Biological Sciences, University of Lancaster, England.
Huckerby
T N
TN
Nieduszynski
I A
IA
D000118:Acetylglucosaminidase / Q000378:metabolism*; D000818:Animals; D001407:Bacillus / Q000201:enzymology; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002358:Cartilage, Articular / Q000737:chemistry*; D002417:Cattle; D002852:Chromatography, Ion Exchange; D006868:Hydrolysis; D007632:Keratan Sulfate / Q000737:chemistry* / Q000302:isolation & purification; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000032:analysis; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D012794:Sialic Acids / Q000032:analysis
D009005:Monosaccharides; D009844:Oligosaccharides; D012794:Sialic Acids; D007632:Keratan Sulfate; C086950:keratanase II; D000118:Acetylglucosaminidase