The transfer of liquid hydrocarbons into water is accompanied by a large decrease in volume at 25 °C and atmospheric pressure, with typical values for ΔV°tr of — 2.0 ml mol methylene−1. Considering the large amount of apolar surface that is exposed when a globular protein unfolds, the hydrocarbon transfer results imply that the change in volume accompanying the unfolding process (ΔV°obs) should be highly negative under these conditions. However, experimental data on the pressure denaturation of proteins typically yield relatively small values of ΔV°obs at atmospheric pressure and 25 °C. We analyze this apparent inconsistency in terms of a simple thermodynamic dissection of the partial molar volume. This approach allows the volume effects that result from solute-solvent interactions to be determined from experimental partial molar volumes. The use of absolute quantities (partial molar volumes) circumvents assumptions associated with the use of results from transfer experiments. An important finding is that hydration of apolar species is less dense than bulk water. This discovery leads to the conclusion that the contribution to ΔV°obs for protein unfolding from the hydration of apolar surfaces is highly positive, contrary to predictions based on transfer data. Further, hydration of polar surfaces makes a positive contribution to ΔV°obs. The large, positive term from the differential hydration of the folded and unfolded states is compensated by the difference in free volume of the protein in the two states. This finding provides a new framework for interpreting pressure effects on macromolecules. The full characterization of a macromolecular system requires knowledge of the effect of pressure on the system. The thermodynamic information obtained from using pressure as a perturbation is a volume change for the particular reaction being studied. The observed volume change, ΔV°obs, for protein unfolding may provide insight into the mechanisms that determine the three-dimensional structure of the folded state. Pressure denaturation experiments have been demonstrated for a number of proteins, including ribonuclease A (Gill & Glogovsky, 1965; Brandts et al., 1970), chymotrypsinogen (Hawley, 1971), metmyoglobin (Zipp & Kauzmann, 1973), and, more recently, lysozyme (Samarasinghe et al., 1992) and staphylococcal nuclease (Royer et al., 1993).
Studies on the Aqueous Solutions of Guanidinium Salts. VII. Activity Coefficients and Partial Molar Volumes for Guanidinium Salts with Hydrophobic Counter Anions in Aqueous Solutions
