Desolvation is a likely origin of robust enthalpic barriers to protein folding

Experimental data front global analyses of temperature (T) and denaturant dependence of the folding rates of small proteins led to an intrinsic enthalpic folding barrier hypothesis: to a good approximation, the T-dependence of folding rate under constant native stability conditions is Arrhenius. Furthermore, for a given protein, the slope of isostability folding rate versus 1/T is essentially independent of native stability. This hypothesis implies a simple relationship between chevron and Eyring plots of folding that is easily discernible when both sets of rates are expressed as functions of native stability. Using experimental data in the literature, we verify the predicted chevron-Eyring relationship for 14 proteins and determine their intrinsic enthalpic folding barriers, which vary approximately from 15 kcal/mol to 40 kcal/mol for different proteins. These enthalpic barriers do not appear to correlate with folding rates, but they exhibit correlation with equilibrium unfolding enthalpy at room temperature. Intrinsic enthalpic barriers with similarly high magnitudes apply as well to at least two cases of peptide-peptide and peptide-protein association, suggesting that these barriers area hallmark of certain general and fundamental kinetic processes during folding and binding. Using a class of explicit-chain C-alpha protein models with constant elementary enthalpic desolvation barriers between C-alpha positions, we show that small microscopic pairwise desolvation barriers, which are a direct consequence of the particulate nature of water, call act cooperatively to give rise to a significant overall enthalpic barrier to folding, This theoretical finding provides a physical rationalization for the high intrinsic enthalpic barriers in protein folding energetics. Ramifications of entropy-enthalpy compensation in hydrophobic association for the height of enthalpic desolvation barrier are discussed. (c) 2005 Elsevier Ltd, All rights reserved.