Abstract
As a new approach to identify whether, in solution, heparin binds calcium ions by non-specific electrostatic interactions or by formation of a true chelate, comparison is made of 1H n.m.r., 13C n.m.r. and c.d. spectra of calcium and sodium forms of various fractions of heparin with corresponding spectra of related glycosaminoglycans (heparan sulphate and low charge density heparin). The different compounds correspond to a large range of compositions with regard repeating units and sulphate content as shown by potentiometry, conductometry, n.m.r. and c.d. analyses. It is borne out that n.m.r. and c.d. data depend on the nature of the counterion. Furthermore, comparison of n.m.r. and c.d. changes induced by Na +/Ca 2+ counterion exchange look similar for the various compounds except for heparan sulphate. However, it is found that the difference between molar ellipticity values for Ca 2+ and Na + forms is independent of the compound including heparan sulphate. This finding is regarded as showing that no specific Ca 2+ chelating site exists in heparin macromolecules.
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