Markov State Models Underlying the N-Terminal Premodel of TOPK/PBK

Lymphokine-activated killer T-cell-originated protein kinase (TOPK) is a potential target for cancer therapy. To explore the micromechanism, we proposed the N-terminal premodel (NTPM) of the TOPK monomer via homology modeling and molecular dynamic simulations and analyzed the conformational dynamics by Markov state model analysis. The electronegative insert (ENI) motif of the NTPM can be opened with a small probability under wild type, regulated by the so-called "N-C" interaction zone consisting of the N-terminal head, the coil between β3-strand and αC-helix, and the ENI motif. Glutamate substitution at threonine residue 9 or tyrosine residue 74 promotes the closed-open transition, revealing the details of phosphorylation. Allosteric effects induce functionally relevant structural changes, such as increased structural flexibility and active sites, which are thought to be necessary for further activation or binding. These findings provide rational structural templates for designing state-dependent inhibitors and give insight into the molecular regulatory mechanisms of TOPK monomers.