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Possible inflexions in isothermal binding plots derived from equilibrium-dialysis measurements and in equivalent plots obtained by polarimetric measurements accord with the possibility that discrete Ca(2+)-heparin-water complexes/phases may exist, the nature and proportions of which depend on the conditions under which the interaction occurs. Analysis of the plots obtained by polarimetric study of chemically modified heparins suggests, for individual substituents groups, an order of importance of carboxylate greater than N-sulphonate greater than N-acetyl greater than O-sulphate for the Ca(2+)-heparin interaction occurring at [Ca2+]/[heparin disaccharide] ratios of less than 0.5. At higher ratios, transitions occur that eventually lead to the formation of a complex in which the stoichiometry of association is 1 Ca2+ ion/heparin disaccharide unit.

A study of Ca2+-heparin complex-formation by polarimetry