Considering that protein models need to be partially unfolded and refolded to be refined, MD simulations at elevated temperatures may boost this process. This benefit is counteracted by a loss of stability and increased tendency towards thermal denaturation. However, since restraints were applied with respect to the initial model, protein unfolding may be prevented. Thus, improved sampling may be possible with moderately increased temperatures together with restraints. We explored here whether there is an optimal temperature for refinement simulations in the presence of restraints where improved sampling can be achieved without a loss of protein stability. To address this question, refinement runs were tested at different temperatures: 298.15, 320, 340, 360, and 380 K. In these simulations, we used the combined restraint according to Eq. 3.