Effect of High Temperatures on Proteinase K Crystallization Predicted by Molecular Dynamics Simulation

Molecular dynamics simulations revealed notable thermal stability of the precursor cluster (the dimer that forms prior to the crystallization) of proteinase K crystals in the temperature range of 20–60°C. The analysis of the intermolecular contact area in the dimer confirmed that its structure is preserved up to 60°C. This finding led to the hypothesis that this thermophilic protein can be crystallized at a temperature corresponding to its maximum catalytic activity (50–60°C). This theoretical prediction was confirmed experimentally: crystals of proteinase K were successfully grown after incubation at 60°C. These results demonstrate the practical value of the molecular dynamics approach as a tool for predicting crystallization conditions and offer prospects for determining the structure of proteinase K in the functionally relevant conformation, which is in line with current trends in structural biology of thermophilic proteins.