Hydration shell differentiates folded and disordered states of a Trp-cage miniprotein, allowing characterization of structural heterogeneity by wide-line NMR measurements

Hydration properties of folded and unfolded/disordered miniproteins were monitored in frozen solutions by wide-line H-1-NMR. The amount of mobile water as function of T (-80 degrees C -15 degrees C temperature range, wide-line H-1-NMR detects the heterogeneity of protein fold, providing the size of the hydration shell surrounding the accessible conformers at a given temperature. Results indicate that freezing of protein solutions proceeds by the gradual selection of the enthalpically most favored states that also minimize the number of bridging waters