A simple experimental model for hydrophobic interactions in proteins.

The compound N-cyclohexyl-2-pyrrolidone contains a substantial apolar region as well as a peptide bond-like moiety. This solvent, therefore, provides a useful model for protein interiors. Under certain conditions of temperature and salt concentration, cyclohexylpyrrolidone forms a two-phase system with water. This permits partition coefficients and subsequent free energies of transfer of amino acid side chains from cyclohexylpyrrolidone to water to be simply determined. Free energies of transfer measured in this manner for 21 amino acids are found to be substantially less than those obtained from the commonly used ethanol/water solubility model. This suggests less of a contribution of hydrophobic interactions to the stabilization of protein structure than is conventionally assumed.