Skip to main content

Table 2 Kinetic and thermostability vs predicted properties of ALAS2 missense mutantsa

From: Molecular expression, characterization and mechanism of ALAS2 gain-of-function mutants

Parameters

Wild type

R559H

E565D

R572C

S573F

Y586F

Purified SA (U/mg)

93,500

100,000

122,000

121,000

115,000

109,000

Fold–increase

1.00

1.06

1. 30

1.29

1.23

1.17

Vmax

 Succinyl-CoA (U/mg) ± StdDev

93,500±5300

175,000±2190

143,000±7000

136,000±14,100

120,000±6710

146,000±10,100

 Fold-increase

1.00

1.87

1.53

1.45

1.26

1.56

Km

 Succinyl-CoA (μM)

57.1 ± 6.7

46.3 ± 6.5

50.8 ± 2.15

46.7 ± 1.2

41.1 ± 9.2

44.0 ± 5.2

 Fold-decrease

1.00

1.2

1.1

1.2

1.4

1.3

 Hill coeff. (n)

1.6 ± 0.1

1.65 ± 0.08

1.5 ± 0.02

1.7 ± 0.17

1.45 ± 0.11

1.49 ± 0.14

 T1/2 45 °C (min)

4.7 ± 1.4

13.8 ± 3.4

13.6 ± 2.8

11.8 ± 1.5

9.3 ± 0.8

7.5 ± 0.2

Prediction Program (Ref) and Scoreb

PolyPhen-2 (Adzhubei et al. 2010)

 

Benign

Benign

~Damaging

~Damaging

Benign

 Score

 

0.003

0.001

1.000

0.995

0.016

SIFT (Kumar et al. 2009)

 

Tolerated

Tolerated

Deleterious

Deleterious

Tolerated

 Score

 

0.561

0.335

0.000

0.001

0.101

Mutation Assessor (Reva et al. 2011)

 

Low

Neutral

Low

Medium

Neutral

 Score

 

1.32

−0.285

1.94

2.12

0.46

Provean (Choi et al. 2012)

 

Neutral

Neutral

Deleterious

Deleterious

Neutral

 Score

 

−0.45

−0.47

−3.28

−2.75

−0.89

  1. aData are means ±SD for n = 4–5 separate experiments and Vmax, Km, and T1/2 results all show statistically significant differences from the corresponding wild-type values
  2. bData are the values returned by the online prediction servers for each listed program