Pressure perturbation calorimetry measurements on a range of cyclodextrin adamantane, protein ligand (lysozyme-(GlcNac)(3) and ribonuclease-2'CMP) and protein-protein (cytochrome c peroxidase-pseudoazurin) complexes in aqueous solution show consistent reductions in thermal expansibilities compared to the uncomplexed molecules. Thermodynamic data for binding, obtained by titration calorimetry, are also reported. Changes in molar expansibilities can be related to the decrease in solvation during complexation. Although reasonable estimates for numbers of displaced water molecules may be obtained in the case of rigid cyclodextrin-adamantane complexes, protein expansibility data are less easily reconciled. Comparison of data from this wide range of systems indicates that effects are not simply related to changes in solvent-accessible surface area, but may also involve changes in macromolecular dynamics and flexibility. This adds to the growing consensus that understanding thermodynamic parameters associated with noncovalent interactions requires consideration of changes in internal macromolecular fluctuations and dynamics that may not be related to surface area-related solvation effects alone.