Heparan Sulphate Polymers modified by specific chemical desulphation
These heparan sulphates (HS) were prepared by targeted desulphation of a highly-purified preparation of porcine heparan sulphate derived from intestinal mucosa. This material also has a domain structure like that of cell surface HS. Please note that this HS is a different preparation from the GAG-H01 that was used to prepare the HS fractions described above.
Standard chemical methods were used to selectively remove sulphate groups from C2 of iduronic acid (de2S HS), C6 of glucosamine (de6S HS) or the N-sulphate from N-sulphated glucosamine (deNS HS); the deNS HS contains free amino groups due to the loss of N-sulphates. In deNS/Ac HS the free amino groups have been acetylated.
Gel filtration profiles of the modified HS on Superdex S-75 hplc revealed only very minor reductions in molecular size compared to the parental material probably due to the loss of sulphate groups.
With the exception of 6-O desulphated HS the procedures lead to the loss of over 95% of the targeted sulphate groups.
6-O desulphated HS; in this preparation the reduction in 6-O-sulphate groups was 70%. 6-O-sulphates were completely removed from the trisulphated disaccharide (UA,2S – GlcNS,6S) but the targeted desulphation of UA – GlcNAc,6S and UA – GlcNS,6S was incomplete (50% and 70% loss of 6-O-sulphates respectively).
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Disaccharide analysis of selectively desulphated Heparan Sulphates: comparison of parental material with the desulphated preparations.