As an example, "4w52-benzene system" should be read as "the system of benzene bound to T4 L99A started from the 4w52 structure". Preparation and Parametrization of Proteins and Ligands. We prepared the T4 lysozyme protein and ligand structures as in-put structures for MD simulations. The topology and coordinate input files for GROMACS simulations can be found in on GitHub at . We prepared the structure for each protein of interest (4w52, 4w57, and 4w59) using Open-Eye Spruce 14 to add hydrogens and any missing loops. We solvated each protein with TIP3P waters and ions to achieve a concentration of 150 mM. We then parameterized the solvated systems using the Amber ff14SB force field. We assigned partial charges for each ligand of interest -benzene, butylbenzene and hexylbenzene -using OpenEye's AM1-BCC charge engine 16 and parameterized them using Open Force Field version 2.0.0. We combined the solvated protein structures and prepared ligands into native and mixed system complexes as described in the previous subsection to produce a total of nine starting structures (Table ). We used the crystallographic ligand pose from the native structure for each protein-ligand complex. Running molecular dynamics in GRO-MACS. We simulated all protein-ligand complex systems using GROMACS (v.2021.2). Prior to production, we performed energy minimization for up to 1500 steps using steepest descents. We then equilibrated the systems in two phases, using a 20 ps NVT simulation followed by a 5 ns NPT simulation. We ran production simulations for 100 ns per replicate, with a total of 10 repli-cates per system. The MDP files for GRO-MACS simulations can be found on GitHub at .