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Table 4 Results from binding and activity studies on mutated TtSlyD variants

From: Molecular insights into substrate recognition and catalytic mechanism of the chaperone and FKBP peptidyl-prolyl isomerase SlyD

Construct

Mutation

N1

KD1 (μM)

ΔH1 (kcal/mol)

–T · ΔS1 (kcal/mol)

N2

KD2 (μM)

ΔH2 (kcal/mol)

–T · ΔS2 (kcal/mol)

Activity (kcat/KM 106 M/s)

Relative activity (%)

TtSlyDFL

Wild type

0.96 ± 0.03

0.113 ± 0.012

–10.0 ± 0.11

0.7

0.96 ± 0.02

2.93 ± 0.12

–4.9 ± 0.22

–2.5

1.47 ± 0.05

100

Y13F

0.85 ± 0.03

0.107 ± 0.011

–10.6 ± 0.14

0.5

0.87 ± 0.02

3.94 ± 0.14

–5.9 ± 0.47

–2.6

1.35 ± 0.17

92

D23A

0.92 ± 0.03

0.075 ± 0.042

–2.0 ± 0.11

1.3

0.88 ± 0.01

1.65 ± 0.15

–6.0 ± 0.11

–2.1

1.15 ± 0.21

78

N35A

0.98 ± 0.02

0.169 ± 0.032

–9.7 ± 0.11

0.6

1.06 ± 0.05

5.18 ± 0.89

–2.5 ± 0.33

–4.6

1.04 ± 0.28

71

I37G

0.93 ± 0.05

0.134 ± 0.038

–9.0 ± 0.11

–0.3

0.91 ± 0.03

3.64 ± 0.33

–6.6 ± 0.37

–0.7

0.73 ± 0.12

50

Y63F

0.81 ± 0.01

0.030 ± 0.003

–11.3 ± 0.11

1.1

0.78 ± 0.02

1.58 ± 0.22

–6.0 ± 0.29

–2.2

1.12 ± 0.11

76

Y63A

0.80 ± 0.01

0.041 ± 0.007

–9.4 ± 0.06

–0.5

0.84 ± 0.02

3.18 ± 0.29

–4.7 ± 0.15

–2.7

0.31 ± 0.19

21

A78G

1.08 ± 0.02

0.262 ± 0.023

–8.2 ± 0.06

–0.6

1.09 ± 0.02

5.24 ± 0.43

–2.8 ± 0.19

–4.3

0.76 ± 0.14

52

Y92A

0.96 ± 0.02

0.254 ± 0.022

–8.7 ± 0.51

–0.1

0.92 ± 0.01

4.33 ± 0.14

–5.3 ± 0.15

–1.9

0.53 ± 0.12

36

Q94A

1.03 ± 0.03

0.288 ± 0.042

–8.6 ± 0.10

–0.2

0.93 ± 0.02

5.98 ± 0.44

–4.3 ± 0.38

–2.8

1.44 ± 0.49

98

M96A

1.00 ± 0.01

0.129 ± 0.009

–9.7 ± 0.04

0.4

1.01 ± 0.01

3.86 ± 0.21

–4.4 ± 0.13

–2.9

0.58 ± 0.15

39

H119A

0.87 ± 0.03

0.060 ± 0.012

–11.1 ± 0.05

0.7

0.82 ± 0.02

2.22 ± 0.28

–6.2 ± 0.47

–2.2

1.23 ± 0.02

84

F128A

0.88 ± 0.02

0.215 ± 0.022

–9.7 ± 0.36

1.2

0.89 ± 0.02

1.62 ± 0.11

–8.0 ± 0.52

–1.2

0.52 ± 0.16

35

TtSlyDΔIF

Wild type

    

0.88 ± 0.02

12.23 ± 0.24

–5.9 ± 0.11

0.4

0.85 ± 0.01

58

  1. The affinities and thermodynamic parameters were determined at 20 °C for the wild type S2 peptide using isothermal titration calorimetry, while the activities were determined for the suc-ALPF-pNA tetrapeptide using nuclear magnetic resonance spectroscopy. TtSlyD ΔIF TtSlyD constructs with the insert-in-flap domain replaced by the flap loop from human FKBP12, TtSlyD FL full-length TtSlyD
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BMC Biology

ISSN: 1741-7007

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