Nucleotide-dependent dynamics of KIF-14 conserved structural domains in kinesin

Atomistic simulations of kinesin molecular motor proteins reveal the interplay between structure and dynamics to power movement. Here we analyze the kinesin-3 molecular motor KIF14 using all-atom molecular dynamics simulations of the catalytic core bound to tubulin in the Apo, ADP, ADP+Pi, and ATP kinesin ligand-binding states, initialized from high-quality cryo-EM structures (Benoit et al., 2021). Each complex contains explicit water (~285k atoms) and is simulated for 100 ns and with multiple replicates. We compare the intrinsic dynamics and local atomic environments across states, focusing on hallmark motifs—Switch I, Switch II, the P-loop, and the α4 helix—and on the state-dependent emergence of the R604–E643 salt bridge. To quantify coordinated motion, we compute amino-acid distance metrics (e.g., Cα–Cα and side-chain centroid distances) within and between subdomains, generating state-resolved distance maps that report on open vs. closed states and pathway-critical contacts. Together, these data provide insight into a conserved "parts list" for motor function and establish an atomic-scale baseline for understanding how ATP hydrolysis is transduced into mechanical motion, both within KIF14 and in comparisons across the kinesin superfamily.