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 7 Effect of single amino acid substitution on MAPK1 and MAPK3 variants stability and catalysis

	^aTertiary structure changes		^bThree-state unfolding		^cTwo-state unfolding		^dΔTm (°C)		^ekcat/K_m ratio	^dΔTmax (°C)	^dΔEa (kcal /mol)
	NP	P	NP	P	NP	P	NP	P
MAPK1
Wild type	–	–	No	No	Yes	Yes	–	–	–	–	–
Group I and II
E81K	Yes	Yes	Yes	Yes	No	No	0	− 0.9	0.39	5	− 3.05
R135K	No	No	Yes	No	No	Yes	0.1	0.1	3.75	5	− 2.89
D162G	Yes	Yes	No	Yes	Yes	No	− 0.5	5.0	5.33 × 10^–4	10	− 3.01
R191H	Yes	Yes	Yes	No	No	Yes	3.0	3.1	2.98 × 10^–4	10	− 1.86
Y316F	Yes	Yes	No	No	Yes	Yes	0.1	0.1	3.57	7	− 4.21
P319S	Yes	Yes	Yes	Yes	No	No	− 4.0	0.1	6.97 × 10^–2	5	− 2.39
E322V	Yes	Yes	Yes	Yes	No	No	2.1	1.0	1.11	7	− 5.96
Group III
E33Q	Yes	Yes	No	Yes	Yes	No	− 0.9	4.0	4.16 × 10^–1	− 5	1.15
L121I	Yes	Yes	No	No	Yes	Yes	0	7.0	8.28 × 10^–3	5	− 0.98
L200F	Yes	Yes	No	No	Yes	Yes	− 1.0	1.1	6.16 × 10^–3	0	− 4.16
D235V	Yes	Yes	No	No	Yes	Yes	− 2.9	6.1	6.40 × 10^–2	7	− 1.31
MAPK3
Wild type	–	–	No	No	Yes	Yes	–	–	–	–	–
Group I and II
E98K	No	No	Yes	Yes	No	No	6.0	0	1.66	− 12	1.60
R152W	Yes	Yes	Yes	Yes	No	No	3.0	2.9	2.46 × 10^–1	3	1.29
P336Q	Yes	Yes	Yes	Yes	No	No	− 2.0	− 0.1	3.5 × 10^–1	0	1.98
E339V	No	No	Yes	Yes	No	No	0.1	1.0	5.69 × 10^–1	− 7	0.53
Group III
I73M	Yes	Yes	Yes	Yes	No	No	− 1.0	4.9	4.90 × 10^–1	3	0.55
Q79H	Yes	Yes	Yes	Yes	No	No	5.5	0	7.69 × 10^–2	− 12	4.33
A160T	Yes	Yes	Yes	Yes	No	No	− 3.9	− 3.0	9.23 × 10^–1	− 12	5.04
T198I	Yes	Yes	Yes	Yes	No	No	20.5	0.9	5.046	− 12	3.88
E214D	Yes	Yes	Yes	Yes	No	No	14.3	8.9	9.23 × 10^–2	0	4.47
L281I	Yes	Yes	Yes	Yes	No	No	0.1	0	6.46 × 10^–1	− 7	5.92
V290A	Yes	Yes	Yes	Yes	No	No	1.0	8.9	1.38 × 10^–1	− 17	8.42
R359W	Yes	Yes	Yes	Yes	No	No	− 2.9	9.0	7.08 × 10^–1	− 2	2.09
E362K	Yes	Yes	Yes	Yes	No	No	− 1.9	− 1.1	2.61 × 10^–1	− 12	6.82

^aObserved by comparison of the near-UV CD spectra of the variants with that of the wild type. NP and P indicate the non-phosphorylated and phosphorylate conformation of the proteins, respectively
^bGdmCl equilibrium unfolding fitted to Eq. (4)
^cGdmCl equilibrium unfolding fitted to Eq. (6). T_m is the melting temperature; T_max is the temperature for maximal activity; E_a is the activation energy obtained from Eq. (1)
^dΔcorresponds to the difference between the variant and the wild type
^ekcat/K_m ratio = kcat/K_mvariant/kcat/K_mwild type

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