Table 1.

Kinetic and thermodynamic parameters obtained from the analysis of pseudo-first order binding kinetics between site-directed variants of N-SH2 and Gab2_608–620.

	kon (μM−1 s−1)	koff (s−1)	KD (nM)	Activation Free Energy ∆∆G# (kcal mol−1)	Equilibrium Free Energy ∆∆Geq (kcal mol−1)
WT	19.0 ± 0.5	1.85 ± 0.01	100 ± 5
I11V	22.2 ± 1.5	1.81 ± 0.01	80 ± 6	−0.09 ± 0.02	−0.10 ± 0.01
T12S	21.2 ± 1.4	1.73 ± 0.01	80 ± 6	−0.06 ± 0.01	−0.10 ± 0.02
V14A	17.9 ± 0.6	2.81 ± 0.01	160 ± 5	0.03 ± 0.01	0.27 ± 0.03
L19A	13.8 ± 0.5	2.39 ± 0.01	170 ± 6	0.18 ± 0.02	0.32 ± 0.03
T22S	19.2 ± 0.8	1.93 ± 0.01	100 ± 4	−0.01 ± 0.01	0.02 ± 0.01
V25A	20.2 ± 0.9	2.04 ± 0.07	100 ± 6	−0.04 ± 0.01	0.02 ± 0.01
A31G	23.5 ± 1.1	1.62 ± 0.01	70 ± 3	−0.12 ± 0.02	−0.19 ± 0.02
T42S	28.6 ± 1.1	0.14 ± 0.01	5.0 ± 0.5	−0.23 ± 0.02	−1.69 ± 0.10
V45A	14.1 ± 0.7	3.23 ± 0.01	230 ± 11	0.17 ± 0.10	0.48 ± 0.05
A50G	23.0 ± 1.0	1.63 ± 0.07	70 ± 4	−0.11 ± 0.01	−0.18 ± 0.02
T52S	18.7 ± 1.0	0.20 ± 0.01	10 ± 1	0.01 ± 0.01	−1.24 ± 0.10
I54V	17.0 ± 0.5	2.53 ± 0.20	150 ± 13	0.06 ± 0.03	0.24 ± 0.02
I56V	26.0 ± 1.1	0.68 ± 0.01	30 ± 1	−0.18 ± 0.02	−0.74 ± 0.07
T59S	19.1 ± 0.9	2.30 ± 0.01	120 ± 6	0.00 ± 0.01	0.12 ± 0.01
L65A	14.8 ± 1.5	12.70 ± 0.10	860 ± 90	0.14 ± 0.05	1.22 ± 0.10
A72G	14.7 ± 0.9	2.39 ± 0.01	170 ± 10	0.14 ± 0.02	0.29 ± 0.03
T73S	19.7 ± 0.8	2.09 ± 0.02	110 ± 4	−0.02 ± 0.01	0.05 ± 0.01
L74A	15.6 ± 1.9	3.23 ± 0.01	210 ± 25	0.11 ± 0.05	0.43 ± 0.04
A75G	20.7 ± 2.1	2.35 ± 0.02	110 ± 10	−0.05 ± 0.02	0.09 ± 0.01
V78A	14.5 ± 1.7	2.73 ± 0.40	190 ± 35	0.15 ± 0.04	0.37 ± 0.03
L88A	14.0 ± 0.9	10.30 ± 0.05	730 ± 50	0.17 ± 0.05	1.13 ± 0.10
V95A	18.9 ± 1.7	2.44 ± 0.01	130 ± 12	0.00 ± 0.02	0.16 ± 0.01
I96V	20.5 ± 0.8	2.06 ± 0.01	100 ± 4	−0.04 ± 0.02	0.02 ± 0.01