Interaction between proteins and salt solutions. IV. Enrichment of salts in collagen during denaturation

Using 14 C‐labeled KSCN and 36 Cl‐labeled KC1, we have determined the selective removal of ions and water by fibrous, crosslinked collagen both below and above the denaturation temperatures at several molarities under or near isoelectric conditions. The results for KSCN show there is an increase in ion binding at the denaturation temperature. These data, in conjunction with date from the literature, have been used to evaluate binding constants and mole fractions of available binding sites, as well as the free energy of binding various salts to collagen and gelatin. The values so obtained correlate satisfactorily with similar quantities obtained from the experimental dependence of the shrinkage temperature of collagen on salt concentration using a simple theory for the melting point depression which includes the effect of binding in the denatured state. Salting‐out agents show negligible binding to the protein, and this confirms the earlier finding that the interaction between these agents and the protein can be approximately described by conventional polymer–diluent theories which do not consider ion binding. Also, an analysis of the role of unequal anion and cation binding is presented.