*3.10. MD Simulation Studies*

MD simulation of edG alone and edG-HS complex was performed using GROMACS version 2018-2 [72]. The topology of protein structure was generated using the GROMOS96 43a2 force-field [73]. The topology of HS was generated using the PRODRG server [74]. After generating the topology of docked complex, salvated through the SPC/E water model and appropriate Na<sup>+</sup> and Cl<sup>−</sup> ions were added for neutralization [75]. The system's energy was minimized using the combined steepest descent algorithm, a convergence criterion of 0.005 kcal/mol. The equilibrium condition was developed by NVT (constant volume) and NPT (constant pressure) at a 100 ps time scale. Berendsen weak coupling method was used to maintain the system's temperature at 298 K, and Parrinello–Rahman barostat was used to adjust the pressure of 1 bar in the equilibrium condition. The final conformational production stage of the time scale of 50 ns was generated using the LINCS algorithm, and generated trajectories were analyzed to know the behaviour of the complex in the explicit water milieu. The root-mean-square deviations (RMSD), root-mean square fluctuation (RMSF), the radius of gyration (Rg), solvent-accessible surface area (SASA), Hydrogen bonds, the free energy landscape of complex, and conformational changes were analyzed.
