Ligand unbinding mechanisms and kinetics for T4 lysozyme mutants from τRAMD simulations

The protein-ligand residence time, τ, influences molecular function in biological networks and has been recognized as an important determinant of drug efficacy. To predict τ, computational methods must overcome the problem that τ often exceeds the timescales accessible to conventional molecular dynamics (MD) simulation. Here, we apply the τ-Random Acceleration Molecular Dynamics (τRAMD) method to a set of kinetically characterized complexes of T4 lysozyme mutants with small, engineered binding cavities. τRAMD yields relative ligand dissociation rates in good accordance with experiments across this diverse set of complexes that differ with regard to measurement temperature, ligand identity, protein mutation and binding cavity. τRAMD thereby allows a comprehensive characterization of the ligand egress routes and determinants of τ. Although ligand dissociation by multiple egress routes is observed, we find that egress via the predominant route determines the value of τ. We also find that the presence of a greater number of metastable states along egress pathways leads to slower protein-ligand dissociation. These physical insights could be exploited in the rational optimization of the kinetic properties of drug candidates.
Graphical abstract Image 1
Highlights • Relative residence times are computed for T4 lysozyme mutant-ligand complexes. • τ-Random Acceleration Molecular Dynamics provide efficient sampling of unbinding. • Computed dissociation rates show good agreement with all available measured values. • Ligand egress via the predominant route determines the value of the residence time. • The presence of metastable states along egress pathways slows down dissociation.