TABLE 1.

Comparison of fitness metric-based dissociation constant calculations with published experimentally determined dissociation constants

Variant	In vitro binding	Deep sequencing data
	Kd,i/Kd,wt	Kd,i/Kd,wt	Fitness metric
WT	1.00 ± 0.10	1.00	0
R146A	1.07 ± 0.10	1.08 ± 0.02	−0.074
E155A	0.79 ± 0.09	0.97 ± 0.02	0.034
T156A	0.79 ± 0.09	1.04 ± 0.04	−0.037
T159A	1.00 ± 0.16	1.06 ± 0.03	−0.053
Y161A	0.93 ± 0.43	0.88 ± 0.04	0.116
N176A	0.79 ± 0.09	0.81 ± 0.03	0.199
E210A	1.14 ± 0.16	1.05 ± 0.02	−0.052
E16A	1.14 ± 0.11	1.30 ± 0.08	−0.266
T81A	1.21 ± 0.65	1.19 ± 0.03	−0.168
T158A	0.93 ± 0.16	0.98 ± 0.03	0.020
Y166A	1.00 ± 0.16	1.10 ± 0.02	−0.094
R39A	2.14 ± 0.32	0.84 ± 0.06	0.165
T153A	1.43 ± 0.30	1.48 ± 0.03	−0.403
R79A	17.9 ± 3.1	2.62 ± 0.25	−1.058
H83A	7.1 ± 1.5	2.14 ± 0.14	−0.826
Y148A	20.7 ± 4.5	1.69 ± 0.16	−0.546
N150A	5.7 ± 0.8	1.89 ± 0.08	−0.670
T81K	No bindinga	4.25 ± 0.88	−1.664
T81H	1.51 ± 0.43a	2.55 ± 0.24	−1.025
I152M	0.68 ± 0.23a	0.67 ± 0.11	0.359
I152P	1.36 ± 0.46a	2.12 ± 0.06	−0.806

^a Reported numbers are experimental relative EC₅₀ values.