The populations calculated from this MSM agree well with values obtained from calculated absolute binding free energies for the different binding modes. We performed absolute binding free energy calculations from the two non-symmetric binding modes (see Section 2.3.5. The binding free energy for the crystallographic binding mode was -7.1±0.2 k B T , while the non-crystallographic binding mode was calculated to be -6.5±0.2 k B T . The populations estimated from the MSM would translate to a free energy difference of approximately 0.41 k B T , which is within error of the difference in absolute binding free energies, 0.6 ± 0.3 k B T .