Table 3.

Free energy estimates, the resulting average free energies, and the relative free energies for all processes defined in Fig. 1 and compared to experimentally determined values

	Forward	Backward	Averagea	Hysteresisb
	−0.4 ± 1.5	−1.0 ± 1.8	−0.7	0.6
	10.6 ± 1.5	19.1 ± 1.7	12.2	8.5
	3.6 ± 2.1	6.8 ± 1.9	4.7	3.2
	35.8 ± 3.0	36.0 ± 3.5	35.9	0.2
	36.8 ± 2.0	40.2 ± 2.7	38.0	3.4
	50.5 ± 4.3	41.3 ± 2.5	42.9	9.2
	20.7 ± 11.4	13.4 ± 8.8	12.5

			Averagea	Experimental
Approach 1
			12.9	0.8
			5.4	−6.9
Approach 2
			−7.5	−7.7
Approach 3
			2.1	7.4
			7.0	0.3
Approach 4
(WT,F483A)			5.0	−7.1

^aAverage determined according to Eq. 3

^bHysteresis, defined as the difference between performing a process in the forward and backward directions, yields information on the convergence of the two independent calculations