Abstract
Many proteins denature when they are transferred to concentrated urea solutions. Three mechanisms for urea's denaturing ability have been proposed: (i) direct binding to polar parts of the protein surface, (ii) direct binding to nonpolar parts of the protein surface, and (iii) an indirect effect mediated by modifications of the bulk water properties. The disentanglement of these three processes ...
Abstract
Many proteins denature when they are transferred to concentrated urea solutions. Three mechanisms for urea's denaturing ability have been proposed: (i) direct binding to polar parts of the protein surface, (ii) direct binding to nonpolar parts of the protein surface, and (iii) an indirect effect mediated by modifications of the bulk water properties. The disentanglement of these three processes has been the goal of many experimental and computational studies, yet there is no final agreement on the relative importance of the three contributions. The separation of the two direct mechanisms, albeit conceptually clear, is difficult in experimental studies and in simulations depends subtly on how the discrimination between polar and nonpolar groups is accomplished. Indirect effects, embodied in the change of solution activity as urea is added, are rarely monitored in urea/peptide simulations and thus have remained elusive in numerical studies. In this paper we establish a rigorous separation of all three contributions to the solvation thermodynamics of stretched peptide chains. We contrast this scenario with two commonly used model systems: the air/water interface and the interface between water and a hydrophobic alkane self-assembled monolayer. Together with bulk thermodynamic properties of urea/water mixed solvents, a complete thermodynamic description of the urea/water/peptide system is obtained: urea avoids the air/water interface but readily adsorbs at the oil-water interface and at hydrophobic as well as hydrophilic peptide chains, in accordance with experimental results. Simple thermodynamic arguments show that the indirect contribution to urea's denaturing capability is negligibly small, although urea strongly changes the water bulk properties as judged by the number of hydrogen bonds formed. Urea's tendency to bind to proteins is correctly reproduced with several force field combinations, but the quantitative binding strength as well as the relative importance of direct and indirect effects vary drastically between different force fields used for urea and the peptides.
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