Observations on the origin of the non-linear van't Hoff behaviour of polypeptides in hydrophobic environments.

In this paper we describe a general procedure to determine the thermodynamic parameters associated with the interaction of polypeptides or proteins with immobilised lipophilic compounds such as non-polar n-octyl groups. To this end, the binding behaviour of an all L-alpha-polypeptide, 1, and its retro-inverso-isomer, 2, has been investigated with an n-octylsilica and water-organic solvent mixture containing different percentages of acetonitrile or methanol over the temperature range of 278-338 K. The results confirm that non-linear van'ts Hoff plots occur with this pair of polypeptide isomers, depending on the solvent composition. These findings are consistent with the changes in the thermodynamic parameters, enthalpy of association, delta Hoassoc,i, entropy of association, delta Soassoc,i, and heat capacity, delta Cop,i, all having significant temperature dependencies. Theoretical relationship linking the changes in the delta Hoassoc,i, delta Soassoc,i and delta Cop,i values of these polypeptide-non-polar ligate systems, as a function of temperature, T, have been validated. Significant differences were observed in the magnitudes of these thermodynamic quantities when acetonitrile or methanol was employed as the organic solvent. The origin of these solvent-dependent effects can be attributed to the hydrogen-bonding propensity of the respective solvent. Involvement of enthalpy-entropy compensation effects associated with the interaction of these polypeptides with the hydrophobic ligates has also been documented. Analysis of empirical extra-thermodynamic relationships associated with molecular structural properties of these polypeptides, such as the slope term, S, derived from the plots of the logarithmic capacity factor, log k'i, of these polypeptides vs. the volume fraction of the organic solvent, [symbol: see text] as a function of temperature, T, has also revealed similar correlations in terms of the interactive behaviour of polypeptides 1 and 2 under these experimental conditions. These findings provide an extended thermodynamic and extra-thermodynamic framework to examine the solvational, conformational and other equilibrium processes that polypeptides (or proteins) can undergo in the presence of n-alkylsilicas or other classes of immobilised hydrophobic surfaces. The experimental approach utilised in this study with these topologically similar polypeptides thus represents a generic procedure to explore the behaviour of polypeptides or proteins in non-polar environments in terms of their molecular properties and the associated linear free energy relationships that determine their interactive behaviour.