Conformational degeneracy restricts the effective information content of heparan sulfate

Abstract

The linear, sulfated polysaccharide heparan sulfate occupies a pivotal position in intercellular signalling events, interacting with numerous proteins on the cell surface and in the extracellular matrix. Its complex sequences suggest high potential information content but, despite extensive efforts, a clear relationship between its substitution pattern and biological activity remains elusive. This results from technical limitations, compounded by attempts to correlate substitution pattern directly with activity without considering other conformational factors. For a series of systematically modified analogues of heparan sulfate, the relationship between substitution pattern and experimental ¹³C NMR chemical shifts, which act as reporters of the presence of conformational change, particularly around the glycosidic linkages, was explored through chemometric analysis. From analysis of the experimental data it was evident that wide linkage variation arose from O-sulfation in iduronate and N-sulfation in glucosamine residues but, their effects were distinct, while 6-O-sulfation had much less impact. Models of saccharide sequences showed that the maximum spread of variation in glycosidic linkages occurred before maximum sequence diversity and revealed a highly degenerate system: a fraction of possible sequences is sufficient to provide diverse backbone conformations to satisfy particular protein binding requirements. The unique information content potentially available in HS sequences, defined ultimately by conformation, is vastly inferior to the potential sequence diversity.