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Table 3 Kinetic parameters of wild-type and mutant variants of SK and AK1

From: The role of the C8 proton of ATP in the catalysis of shikimate kinase and adenylate kinase

Enzyme

Km (mM)

v max

Kcat (s -1)

Kcat/Km (s-1 M -1)

1. SK WT ATP (M-M)

6.533 6.53 ± 3.25

0.0484 233.8 ± 60.58

4.841 × 103

8.749 × 102

2. SK WT d-ATP (M-M)

12.74 19.26 ± 7.00

0.0503 333.4 ± 89.2

5.027 × 103

3.946 × 102

3. SK-T17I ATP (A-S)

Ambig 30.73 ± 2.17

Ambig 43.94 ± 3.87

  

4. SK-T17I d-ATP (M-M)

22.05 ± 10.47 6.22 ± 3.57

0.0333 25.03 ± 7.70

1.331 × 103

60.06

5. SK-K15I ATP (M-M)

4.054 4.13 ± 1.72

0.0057 2.736 ± 0.054

5.664 × 10

1.397 × 10

6. SK-K15I d-ATP (A-S)

1.091 0.754 ± 0.227

0.0023 0.971 ± 0.031

2.369 × 10

2.171 × 10

7. AK1 WT ATP1 (A-S)

1.911

0.00219

6.649 × 103

3.479 × 103

0.2332

141.7

8. AK1 WT d-ATP (M-M)

4.043

0.00250

7.583 × 103

1.875 × 103

8.404

557.7

9. AK1-T23V ATP (M-M)

12.91

0.0148

4.497 × 104

3.483 × 103

12.91

1850

10. AK1-T23V d-ATP (M-M)

10.51

0.00963

2.919 × 104

2.777 × 103

.22

1062

11. AK1-R128A ATP

Ambig

Ambig 4.597

Ambig

 
 

59.90

 

12. AK1-R128A d-ATP

Ambig

Ambig

  

13.36

1.299

13. AK1-T39V ATP (A-S)

5.485

0.00398

1.989 × 103

3.626 × 102

1.191

29.57

14. AK1-T39V d-ATP (M-M)

3.235

0.00159

7.972 × 102

2.464 × 102

4.040

39.21

15. AK1-R97A ATP (A-S)

0.1226

0.00131 2.532

6.527 × 102

5.323 × 103

0.0002

 

16. AK1-R97A d-ATP (M-M)

Ambig

0.0003 4.388

1.605 × 102

 
 

0.4596

 
  1. The kinetic parameters of wild type SK, SK-T17I, SK-K15I, wild type AK1, AK1-T23I, AK1-T39I AK1-R97A and AK1-R128A. The datasets for each enzyme comprise values for the kinetic constants calculated using the non-linear regression analysis to calculate the Kcat (upper value), and either the best-fit data, as outlined by the GraphPad Prism5 for either the Michaelis-Menton (M-M) or Allosteric sigmoidal (A-S) fit (lower value). Ambig = Ambigous.