Table 1

Alanine scan of LEXA-HSF1

Construct

Relative luciferase


C

SD

HS

SD


LEXA-HSF1

0.50

0.1

100

13.5

LEXA-HSF1(F)

0.32

0.1

100

6.3

T97A

0.83

0.3

122

5.0

T120A/S121A(F)

0.59

0.1

115

4.7

S123A/T124A

0.65

0.6

95

6.6

S127A

0.50

0.0

84

5.1

S136A/T138A(F)

0.23

0.1

112

11.6

T142A(F)

0.22

0.0

83

11.0

S156A(F)

0.29

0.0

102

6.1

S(174/195/199)A

0.30

0.0

105

8.2

S216A

0.52

0.1

77

10.5

S218A(F)

0.31

0.0

117

9.5

S221A(F)

0.64

0.4

70

8.3

Y225A/S(226/230)A(F)

0.39

0.0

92

5.8

S(237/241/244)A/Y240A

0.54

0.1

126

6.4

Y247A/S(248/249)A(F)

0.72

0.1

127

2.2

S(250/251)A/Y253(F)

1.21

0.3

99

16.6

S(260/261)A

0.84

0.2

119

19.1

S266A/T269A(F)

0.95

0.3

83

11.8

S275A

1.05

0.3

75

5.5

S279A

0.51

0.0

103

24.7

S290A

0.32

0.0

101

13.5

S291A

1.43

0.4

80

4.1

S292A

1.47

0.6

84

3.1

S303A

0.97

0.3

81

13.9

S307A(F)

0.36

0.0

148

33.7

S(303/307)A(F)

1.27

0.5

249

16.9

S314A(F)

0.34

0.0

131

18.3

S(319/320)A/T323A

0.83

0.2

78

10.8

S326A/T328A

2.15

1.0

46

1.9

S333A(F)

0.37

0.0

103

9.3

S338A(F)

0.51

0.1

88

10.1

S344A(F)

0.47

0.1

101

13.4

T346A(F)

0.60

0.1

108

15.0

T349A(F)

0.35

0.1

95

12.6

T(355/357)A(F)

0.34

0.0

129

12.6

S(363/368)A/T(367/369)A(F)

1.05

0.6

92

14.4

S375A

2.13

1.9

73

13.6

S385A(F)

0.57

0.1

121

5.7

S393A

1.38

0.6

83

14.9

T400A/S(403/404)A(F)

0.57

0.2

63

2.9

T411A/S412A(F)

0.57

0.0

85

6.3

S(419/421)A/T423A(F)

0.48

0.0

101

6.7

S428A

0.88

0.1

75

9.6

S(434/435/438)A

1.39

0.1

81

12.9

S444A

1.27

0.2

88

13.2

S(457/458/461)A(F)

0.49

0.1

7

0.3

Y468A/T469A

2.12

1.9

103

16.6

S(480/485)A/T483A

0.47

0.1

80

4.0

S(498/501)A/Y499A

1.08

0.1

80

9.2

T511A/S513A(F)

0.44

0.1

22

1.0

T516A/S518A(F)

0.87

0.1

78

8.1

T527A/S529A

1.14

0.2

91

4.6

S121A

1.73

0.2

106

12.1

S230A

1.64

0.3

101

22.7

S319A

0.60

0.1

105

14.3

S320A

0.36

0.1

91

12.8

T323A

1.97

0.5

102

17.6

S326A

1.66

0.3

40

8.1

S419A

1.67

0.2

104

17.8


C: not-heated; SD: standard deviation; HS: heat-treated. Transactivation assays were carried out as discussed in Methods and Results. (F) refers to the presence of an amino-terminal FLAG tag (see Methods). The activities of FLAG-tagged mutants are expressed relative to the activity of FLAG-LEXA-HSF1, and those of not-FLAG-tagged mutants to the activity of LEXA-HSF1.

Guettouche et al. BMC Biochemistry 2005 6:4   doi:10.1186/1471-2091-6-4

Open Data