| A heparan sulfate proteoglycan was purified from adult bovine brain and its structure characterized. The major heparan sulfate proteoglycan from whole bovine brain had a molecular weight of >200 KDa and a core protein size of 66 KDa. All core proteins from DEAE fractions were identical in size and reacted with heparan sulfate proteoglycan stub antibody and an antibody made to a synthetic peptide based on rat glypican, suggesting that the major heparan sulfate proteoglycan in adult bovine brain was glypican. The core proteins had identical peptide maps and the internal sequence of a major peptide showed 37.5% sequence similarity with human glypican 5. The glycosaminoglycan chain size of the three forms of this proteoglycan were identical. The disaccharide composition analysis of the heparan sulfate chains on the three forms of the proteoglycan showed that they differed primarily by degree of sulfation.;A new glycosaminoglycan has been isolated from the giant African snail Achatina filica. This polysaccharide had a uniform repeating disaccharide structure of →4)-alpha-D-GlcNpAc(1 → 4)alpha- L-IdoAp2S (1→. Using heparin lyases prepared from Flavobacterium heparinum and a newly isolated heparin lyase from Bacteroides stercoris, two classes of oligosaccharides were prepared from acharan sulfate and their structures were determined as DeltaUAp2S(1 → [4)-alpha- D-GlcNpAc(1 → 4)-alpha-L-IdoAp2S(1→] n4)-D-GlcNpAcalpha,beta (major form, where n = 0, 1, 2, 3) and DeltaUAp(1 →[4)-alpha-D-GlcNpAc(1 → 4)-alpha-L-IdoAp2S(1→]m-D-GlcNpAcalpha,beta (minor form, where m = 1, 2, 3) by using strong-anion-exchange high performance liquid chromatography, capillary electrophoresis, electrospray ionization mass spectrometry and proton nuclear magnetic resonance spectroscopy.;The action pattern of Streptomyces hyaluronate lyase was explored using high resolution capillary electrophoresis after fluorescent labeling at the reducing end of oligosaccharides by reductive amination. Computer simulation studies gave comparable kinetic profiles suggesting that hyaluronate lyase exhibits a random endolytic action pattern. Interestingly, oligosaccharides of certain size were under-represented in these oligosaccharide mixtures suggesting that linkages at spacings of 10 to 12 saccharide units are somewhat resistant to this enzyme. The cause of this resistance might be the result of secondary or higher order structural features present in the hyaluronic acid. |
