Purification and Characterization of Heparan Sulfate Peptidoglycans from Bovine Liver

Abstract

A method for the isolation and purification of peptidoglycan heparan sulfate (HS) from bovine liver is described. Peptidoglycan HS was purified from bovine liver and its structure was characterized. The purity and identity of this HS was determined following treatment with heparin lyases by using polyacrylamide gel electrophoresis. The disaccharide compositional analysis on the peptidoglycan samples, determined by treatment with a mixture of heparin lyases followed by high-resolution capillary electrophoresis, showed that bovine liver HS was more highly sulfated than the standard porcine intestinal mucosal HS, but it contained no measurable amount of trisulfated disaccharide. One-dimensional 1H-NMR spectroscopy was used to calculate the relative amounts of glucuronic and iduronic acid residues that were present. The presence of peptide chain in this HS was confirmed by amino acid composition analysis and its selective biotinylation and conjugation with streptavidin.