earlier observations that binding mode interconversion is quite slow in the buried lysozyme binding site. Given the timescale of the slowest binding mode transitions observed here, obtaining accurate ligand binding mode populations from brute force MD or even MSMs seems particularly costly in this case. Specifically, to generate an accurate representation of the populations either approach will need to observe multiple transitions between binding modes. Especially in the MD case, this would require simulations which are at least 10 times longer than the 100ns timescale for the slower binding mode interconversion events-equivalent to at least 1µs of simulation. While toluene's symmetry could be used to obtain correct populations without adequately sampling the symmetric ring flip, any new ligand differing by a substitution breaking this symmetry (and there are many such ligands which bind in this site, such as the 3-iodotoluene case considered later in this work ) would require adequate sampling of these previously symmetry-equivalent binding modes. In other words, adequate binding mode population estimates would likely require multiple microseconds of simulation time.