Table 4b.

Effect of mutations on the stability of rRBP as measured by chemical unfolding

Protein	Δf(H2O) (kcal/mole)	m (kcal/mole/M)	Gnd50	δΔG(WT−mutant) (water)	δΔG(WT−mutant) (Gnd50)
rRBP	−4.76 ± 0.16	2.67 ± 0.10	1.79 ± 0.01	—	—
rRBP24Y	−2.99 ± 0.17	2.49 ± 0.13	1.22 ± 0.03	−1.77 ± 0.23	−1.47 ± 0.09
rRBP24F	−3.36 ± 0.33	2.46 ± 0.24	0.37 ± 0.03	−1.40 ± 0.22	−1.09 ± 0.04
rRBP22A/24F	−2.78 ± 0.08	2.86 ± 0.09	0.97 ± 0.01	−1.98 ± 0.18	−2.26 ± 0.06
rRBP91H	−4.77 ± 0.23	2.92 ± 0.14	1.77 ± 0.03	0.01 ± 0.28	0.06 ± 0.08
rRBP105F	−4.44 ± 0.15	2.92 ± 0.10	1.52 ± 0.06	−0.32 ± 0.22	−0.43 ± 0.10
rRBP67L/91H	−4.13 ± 0.26	2.61 ± 0.16	1.65 ± 0.01	−0.45 ± 0.31	−0.37 ± 0.04
rRBP67L/91H/105F	−4.61 ± 0.17	3.16 ± 0.11	1.42 ± 0.01	−0.15 ± 0.23	−0.96 ± 0.02

ΔG values are the free energy difference between the unfolded and folded states of a particlular protein.

m values represent the change in stability with GndHCl.

[GND]₅₀ = is the midpoint of the unfolding transition.

δΔG^mut = ΔG^wt − ΔG^mut

δΔG_GND50 = ([GND]₅₀^wt − [GND]₅₀^mut) * (m^wt + m^mut)/2, the free energy of stability at the midpoint of the unfolding transition.