Table 2.

Solvation free energy (kcal/mol) for growing a water molecule in a solution containing one protein molecule (Factor Xa) and 15951 water molecules. The data in the ΔF (32 states) column are the benchmarks. The ΔF (2 states) column shows the estimates when the 10 ns simulation data at the γ = 1 state and the 100 ns simulation data at the γ = 0 state are UWHAMed. The uncertainties marked with star are the lower limits of the uncertainty. The details of uncertainty evaluation are provided in Supporting Information.

#	ε	ΔF (32 states)	ΔF (2 states)	Overlap (10−6)
1	−22.68	−13.84 ± 0.09	−14.0 ± 0.5*	7.5e + 00
2	−21.01	−12.7 ± 0.17*	−13.3 ± 0.4*	3.4e + 00
3	−19.09	−11.0 ± 0.4*	−11.8 ± 0.4*	2.1e + 00
4	−17.47	−12.7 ± 0.13*	–	~ 0
5	−16.16	−11.40 ± 0.07	−11.9 ± 0.4*	2.0e + 00
6	−14.44	−10.36 ± 0.03	−10.51 ± 0.11	3.7e + 02
7	−12.76	−11.43 ± 0.08	−10.7 ± 0.6*	3.2e + 00
8	−11.11	−9.033 ± 0.014	−9.00 ± 0.15	9.1e + 01
9	−9.63	−8.782 ± 0.016	−8.87 ± 0.12	1.4e + 02
10	–7.83	−9.095 ± 0.013	−9.06 ± 0.07	4.9e + 02
11	−6.40	−7.774 ± 0.017	−7.67 ± 0.13	8.1e + 01
12	−5.78	−8.895 ± 0.012	−8.92 ± 0.09	1.6e + 02
13	−4.74	−5.765 ± 0.011	−5.85 ± 0.08	1.3e + 02
14	−4.19	−5.66 ± 0.02	−5.82 ± 0.14	8.2e + 01
15	−2.08	−9.33 ± 0.06	−9.26 ± 0.09	1.2e + 03
16	−0.015	−6.013 ± 0.010	−6.02 ± 0.08	2.9e + 02
17	1.04	−4.731 ± 0.015	−4.61 ± 0.12	9.9e + 01
18	3.14	−6.200 ± 0.013	−6.19 ± 0.13	1.6e + 02
19	5.29	−4.578 ± 0.014	−4.46 ± 0.08	5.9e + 02
20	7.04	−11.0 ± 0.12*	–	~ 0
21	8.94	−8.60 ± 0.14	−7.7 ± 0.6*	3.5e + 00