Table 7.

Calculated relative binding free-energies and standard errors (obtained with MBAR in kJ/mol) for the studied perturbations

Transformation	Exp.	Results 1	Results 2	Results 3
Set 1		Conf. 1	Conf. 2
ref → 80		1.8 ± 0.5	−3.6 ± 0.5
81 → 82	10.0	−13.2 ± 0.5	−16.4 ± 0.5
82 → ref		13.3 ± 0.3	16.4 ± 0.3
83 → ref		3.6 ± 0.5	−3.5 ± 0.5
84 → ref		8.3 ± 0.5	8.3 ± 0.6

Set 2 without Wat1/2		2WI7	3FT5	2WI7/3FT5
101 → 100	13.9	−12.2 ± 0.5	2.7 ± 0.5	−12.8 ± 0.5
101 → 105	−0.1	−7.5 ± 0.2	2.7 ± 0.2	−8.4 ± 0.2
101 → 106	2.0	−7.3 ± 0.3	3.8 ± 0.3	−8.7 ± 0.3

Set 2 with Wat1/2		2WI7	3FT5
101 → 100	13.9	11.2 ± 0.9	18.0 ± 0.9
101 → 105	−0.1	−6.2 ± 0.4	3.5 ± 0.4
101 → 106	2.0	−3.7 ± 0.5	5.5 ± 0.5

Set 3		Conf. 1	Conf. 2
10 → ref2		−4.9 ± 0.4	−0.6 ± 0.4
11 → ref2		2.3 ± 0.2
15 → ref3		4.8 ± 0.5	−4.1 ± 0.6
19 → ref1		2.9 ± 0.6
21 → ref3		7.3 ± 0.4	−2.1 ± 0.4
23 → ref2		−6.7 ± 0.5	−13.1 ± 0.6
26 → ref2		3.7 ± 0.4	−12.0 ± 0.4
28 → ref2		1.3 ± 0.4	−1.9 ± 0.4
34 → ref2		−0.2 ± 0.7	−3.7 ± 0.7
61S → ref2		−4.8 ± 0.8
61R → ref2		−19.8 ± 0.4
ref 2 → ref1		−11.5 ± 0.6
ref 3 → ref2		4.5 ± 0.4

Experimental data [23] for the relative energies are also given for the transformations that do not involve any reference ligands