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Table 1 Purification of recombinant wild-type and mutant human ALAS2.

From: Molecular Expression and Characterization of Erythroid-Specific 5-Aminolevulinate Synthase Gain-of-Function Mutations Causing X-Linked Protoporphyria

ALAS2 enzyme

Step

Activity (units)

SA (units/mg)

Purification (-fold)

Yield

Wild type

     
 

Crude extract

775,000

1,520

1

100

 

Amylose affinity

374,000

19,200

13

48

 

Gel filtration

154,000

81,700

54

20

ΔAT

     
 

Crude extract

3,660,000

5,040

1

100

 

Amylose affinity

1,890,000

39,000

8

52

 

Gel filtration

767,000

148,000

29

21

ΔAGTG

     
 

Crude extract

3,660,000

6,130

1

100

 

Amylose affinity

2,590,000

51,500

8

71

 

Gel filtration

896,000

252,000

41

24

ΔG

     
 

Crude extract

1,110,000

1,760

1

100

 

Amylose affinity

769,000

15,300

9

69

 

Gel filtration

620,000

77,500

44

56

p.F557X

     
 

Crude extract

1,840,000

2,290

1

100

 

Amylose affinity

970,000

29,500

13

53

 

Gel filtration

491,000

62,900

27

27

p.Q548X

     
 

Crude extract

4,240,000

7,360

1

100

 

Amylose affinity

2,560,000

65,300

9

60

 

Gel filtration

2,040,000

388,000

53

48

p.Q581X

     
 

Crude extract

567,000

610

1

100

 

Amylose affinity

275,000

7,820

13

48

 

Gel filtration

210,000

69,300

114

37