Molecular profile and mapping of dermatan sulfates from different origins

A method for characterization and molecular profiling of acidic polysaccharides (such as dermatan sulfates) has been developed. A variety of dermatan sulfates, fractionated dermatan sulfates and low molecular weight dermatan sulfates, were examined. First, bacterial lyase-type enzymes (chondroitinase ABC) were used to depolymerize the polysaccharides. Then, mapping of these oligosaccharides (comparable to peptide mapping of proteins) was performed using gradient PAGE and SAX-HPLC. Bands and peaks observed in these maps were identified using oligosaccharide standards of defined chemical structures and physical properties. The resulting map can be used to point to structural differences among these dermatan sulfates regarding their size, charge, degree of sulfation, and contamination. Fine details of fragmentation patterns and absence or presence of contaminants were detected by silver staining of gels. These differences, particularly the content of----4)alpha-IdoA(1----3)- beta-D-GalNAc4S6S(1----sequences (detected using SAX-HPLC as delta UA(1----3)-beta-D-GalNAc4S6S) may play an important role influencing the activity of dermatan sulfates to potentiate HC II inhibition of Factor IIa.