Water at hydrophilic and hydrophobic groups interact differently with proteins. Particularly, hydration properties at hydrophobic groups undergo qualitative changes as temperature decreases below 10 °C. The influence of such interfacial changes on protein dynamics and thermodynamics remains largely unexplored. Here, nanosecond to microsecond protein dynamics and the free energy, enthalpy, and entropy of protein hydration are investigated by in-situ NMR as a function of hydration level and temperature. A crossover at 10 °C in protein dynamics and thermodynamics is revealed. The influence of water at hydrophilic groups shows little temperature dependence, whereas water at hydrophobic groups has stronger effect above 10 °C
