In MM-PBSA, the standard MM energy terms from bonded (bond, angle, and dihedral), electrostatic, and van der Waals, ∆EMM were calculated in the gas phase. The polar component (∆GPB) was evaluated by solving the Poisson-Boltzmann equation linearly, employing a grid spacing of 0.5 Å. The solvent dielectric constant (ε) was set to 80, while the solute's dielectric constant was set to 2. The non-polar component (∆GSASA) was calculated based on the solvent-accessible surface area (SASA), utilizing the constants for the non-polar solvation energy gamma (γ) value of 0.0226 kJ/(mol Å -2 ) and b = 3.84928 kJ/mol . In the g_mmpbsa tool, all energy terms were calculated individually and then combined to assess the contribution of each residue to the overall binding energy.