Folding Pathways of a Knotted Protein with a Realistic Atomistic Force Field

We report on atomistic simulation of the folding of a natively-knotted protein, MJ0366, based on a realistic force field. To the best of our knowledge this is the first reported effort where a realistic force field is used to investigate the folding pathways of a protein with complex native topology. By using the dominant-reaction pathway scheme we collected about 30 successful folding trajectories for the 82-amino acid long trefoil-knotted protein. Despite the dissimilarity of their initial unfolded configuration, these trajectories reach the natively-knotted state through a remarkably similar succession of steps. In particular it is found that knotting occurs essentially through a threading mechanism, involving the passage of the C-terminal through an open region created by the formation of the native -sheet at an earlier stage. The dominance of the knotting by threading mechanism is not observed in MJ0366 folding simulations using simplified, native-centric models. This points to a previously underappreciated role of concerted amino acid interactions, including non-native ones, in aiding the appropriate order of contact formation to achieve knotting. Author Summary: It has been recently observed that the native structure of proteins can contain knots. These are formed during the folding process and are tightened in a specific (i.e. native) location, along the poly-peptide chain. The existence of knots hence implies a high degree coordination of local and global conformational changes, during the folding reaction. In this work we investigate how the knot is formed and what are the dynamical mechanisms which drive the self-entanglement process. To this end, we report on the first atomistically detailed numerical simulation of the folding of a knotted protein, based on a realistic description of the inter-atomic forces. These simulations show that the knot is formed by following a specific sequence of contacts. The comparison of the findings with those based on simplified folding models suggest that the productive succession of contacts is aided by a concerted interplay of amino acid interactions, arguably including non-native ones.