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Table 4 Effect of single amino acid substitution on MAPK3 group III variants stabilitya

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

 

Tm (°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

46.0

1.84 ± 0.12

2.26 ± 0.14

0.93 ± 0.07

1.11 ± 0.08

1.98

2.04

P-WT

47.1

1.72 ± 0.09

2.40 ± 0.12

0.83 ± 0.04

1.22 ± 0.06

2.07

1.97

NP-I73M

45.0

2.38

8.74

3.53

4.60

2.71 ± 0.35

2.74 ± 0.42

3.39 ± 0.37

1.46 ± 0.36

0.88 ± 0.02

3.19 ± 0.08

1.04 ± 0.02

3.16 ± 0.18

P-I73M

46.0

58.0

2.31

6.19

2.29 ± 0.16

2.30 ± 0.33

1.81 ± 0.69

1.20 ± 0.09

1.00 ± 0.03

3.43 ± 0.16

1.91

NP-Q79H

44.1

59.0

2.66

2.99

2.28 ± 0.24

3.19 ± 0.94

1.01 ± 0.20

1.14 ± 0.14

0.83 ± 0.05

2.95 ± 0.12

1.99

P-Q79H

47.1

2.91

2.22

2.38 ± 0.20

3.07 ± 0.46

0.85 ± 0.10

1.33 ± 0.13

0.95 ± 0.03

2.60 ± 0.07

1.79

NP-A160T

42.1

5.52

6.41

1.89 ± 0.09

7.53 ± 0.75

1.78 ± 0.24

0.89 ± 0.05

0.73 ± 0.02

3.59 ± 0.06

2.12

P-A160T

44.1

3.22

2.32

3.28

2.54

3.21 ± 0.68

0.86 ± 0.09

4.31 ± 0.47

1.33 ± 0.14

1.00 ± 0.04

2.70 ± 0.06

0.76 ± 0.01

1.90 ± 0.05

NP-T198I

58.0

75.0

1.45 ± 0.11

2.31 ± 0.09

0.99 ± 0.07

1.01 ± 0.42

1.62

2.29

P-T198I

48.0

3.55

7.84

4.08

3.91

4.37 ± 1.02

2.36 ± 0.72

4.52 ± 0.93

1.31 ± 0.11

0.81 ± 0.03

3.31 ± 0.09

0.90 ± 0.03

3.00 ± 0.04

NP-E214D

48.0

59.0

74.0

1.42

2.58

2.81

3.93

2.40 ± 0.55

0.87 ± 0.29

2.38 ± 0.79

1.64 ± 0.15

0.59 ± 0.06

2.96 ± 0.22

1.18 ± 0.09

2.39 ± 0.03

P-E214D

51.0

61.1

3.96

2.60

2.37

3.25

4.03 ± 0.40

1.05 ± 0.07

2.41 ± 0.40

1.69 ± 0.24

0.99 ± 0.01

2.47 ± 0.04

0.99 ± 0.04

1.93 ± 0.05

NP-L281I

46.1

1.94

5.14

4.38

7.72

2.00 ± 0.22

1.31 ± 0.38

3.59 ± 0.36

2.15 ± 0.29

0.97 ± 0.03

3.90 ± 0.16

1.22 ± 0.02

3.59 ± 0.06

P-L281I

47.1

2.40

2.22

4.72

5.53

2.63 ± 0.63

0.84 ± 0.13

4.50 ± 0.33

1.62 ± 0.23

0.91 ± 0.05

2.64 ± 0.10

1.05 ± 0.01

3.41 ± 0.07

NP-V290A

47.0

2.86

5.24

7.52

2.75

3.05 ± 0.35

1.58 ± 0.33

9.19 ± 1.26

1.14 ± 0.11

0.94 ± 0.02

3.31 ± 0.10

0.82 ± 0.02

2.42 ± 0.06

P-V290A

56.0

1.94

2.54

3.20 ± 0.39

1.97 ± 0.46

0.92 ± 0.16

1.22 ± 0.16

0.98 ± 0.06

2.76 ± 0.10

2.62

NP-R359W

43.1

2.85

3.49

2.51 ± 0.14

3.14 ± 0.66

1.05 ± 0.15

1.04 ± 0.06

0.91 ± 0.04

3.33 ± 0.09

2.40

P-R359W

56.1

2.89

3.49

2.76 ± 0.16

3.17 ± 0.67

1.04 ± 0.14

1.22 ± 0.08

0.91 ± 0.04

3.36 ± 0.04

2.26

NP-E362K

44.1

3.69

3.53

4.11

2.02

4.13 ± 0.81

1.13 ± 0.18

3.83 ± 0.55

0.90 ± 0.10

0.89 ± 0.03

3.11 ± 0.09

1.07 ± 0.02

2.23 ± 0.06

P-E362K

46.0

1.26

5.67

3.51

2.54

2.07 ± 0.18

1.62 ± 0.45

3.42 ± 0.75

0.97 ± 0.16

0.61 ± 0.03

3.51 ± 0.13

1.02 ± 0.04

2.61 ± 0.09

  1. aMelting temperatures (Tm) 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)). ΔGH2O 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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Human Genomics

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