The complex impact of cancer-related missense mutations on the stability and on the biophysical and biochemical properties of MAPK1 and MAPK3 somatic variants

Table 3 Effect of single amino acid substitution on MAPK1 group III variants stability^a

	T_m (°C)	\(\Delta G^{{{\text{H}}_{{2}} {\text{O}}}}\)(kcal/mol)		m (kcal/mol/M)		[GdmCl]_0.5 (M)
		CD ([\(\Theta\)]₂₂₂)	Fluorescence (\(\overline{{\uplambda }}\))	CD ([\(\Theta\)]₂₂₂)	Fluorescence (\(\overline{{\uplambda }}\))	CD ([\(\Theta\)]₂₂₂)	Fluorescence (\(\overline{{\uplambda }}\))
NP-WT	55.0	2.51 ± 0.22	2.79 ± 0.16	0.91 ± 0.09	1.34 ± 0.08	2.76	2.08
P-WT	55.0	2.30 ± 0.16	2.88 ± 0.20	0.97 ± 0.07	1.19 ± 0.09	2.37	2.42
NP-E33Q	54.1	2.44 ± 0.14	3.33 ± 0.31	0.96 ± 0.06	1.97 ± 0.20	2.54	1.69
P-E33Q	59.0	2.22 7.18	2.98 ± 0.24	1.56 ± 0.10 2.86 ± 0.62	1.30 ± 0.11	1.42 ± 0.03 3.43 ± 0.05	2.29
NP-L121I	55.0	2.29 ± 0.14	3.27 ± 0.31	1.00 ± 0.07	1.41 ± 0.14	2.29	2.32
P-L121I	62.0	1.73 ± 0.11	2.82 ± 0.21	1.12 ± 0.07	1.46 ± 0.12	1.55	1.93
NP-L200F	54.0	2.27 ± 0.16	2.88 ± 0.16	0.96 ± 0.07	1.44 ± 0.08	2.36	2.00
P-L200F	56.1	2.15 ± 0.19	3.06 ± 0.30	0.97 ± 0.09	1.72 ± 0.18	2.20	1.77
NP-D235V	52.1	2.46 ± 0.16	2.48 ± 0.19	1.07 ± 0.08	1.15 ± 0.09	2.30	2.15
P-D235V	61.1	1.77 ± 0.11	2.41 ± 0.23	1.07 ± 0.07	1.10 ± 0.12	1.65	2.18

^aMelting temperatures (T_m) were obtained as described in Materials and Methods by continuously monitoring the molar ellipticity at 222 nm every 0.5 °C. GdmCl-induced unfolding equilibrium data were measured as described in Materials and Methods by monitoring the ellipticity at 222 nm ([Θ₂₂₂]) and the fluorescence intensity-averaged emission wavelength (λ¯¯¯λ¯, Eq. (2)). ΔG^H₂O and m values were obtained from Eq. (3); [GdmCl]_0.5 was calculated from Eq. (5). [Θ₂₂₂] and ‾λ data were fitted to Eq. (4) for the 2-state unfolding or to Eq. (6), for the 3-state unfolding. Data are reported as the mean ± SE of the fit. All the measures were performed on the non-phosphorylated (NP-) and phosphorylated (P-) form of the proteins

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