Adsorption of molecules of biological interest onto hydroxyapatite.

Equilibrium and kinetic experiments were conducted to investigate the factors determining the adsorption of salivary macromolecules onto hydroxyapatite. Using amino acids and other small adsorbates, it was determined that the carboxyl attached to the alpha carbon does not appear to adsorb onto HA and the affinities of side-chain carboxyls are much smaller than that of the phosphate group (phosphoserine). Hydroxyl (serine) displays an extremely high affinity, but its adsorption site on HA is different and the number of such sites is much smaller than found for the rest of the functional groups investigated. It is shown that the information obtained from small molecules cannot be readily applied to prediction of the adsorption behavior of salivary macromolecules and polypeptides. The kinetics of adsorption of the salivary phosphopeptide statherin, a polyaspartate, and the salivary prolinerich phosphoprotein PRP3 are consistent with the reversibility of the adsorption process; no conclusion was possible in the case of the protein PRP1. Apparent irreversibility cannot be explained on the basis of multipoint binding or the properties of the carboxyl versus phosphate group; it appears that secondary structure determines to a significant extent the adsorption properties of the macromolecules. Calculation of the thermodynamic molar quantities of adsorption of PRP1, PRP3, and L-ASP onto HA showed that the process is entropically driven. The functional relationship between partial molar entropy and adsorption coverage is similar for the two proteins, but quite different from that for aspartate. Explanations for these results are advanced on the bases of changes in structure configurations and displacement of water from the adsorbate and the adsorbent surface, the second factor being the dominant one in the adsorption of a small molecule such as L-ASP.