Overproduction, Purification and Structural Studies
277
0.14
-
[N,C]]
~A280
1,4
0.12
1.2
0.10
1.0 Z
0
0.00
«l
N
III
()
«
0.8 '---'
-
0.03
6
0.6
o.~
0.4
0.02
0.2
0.00
~l'\x1'
20
40
60
80
100
120
140
160
a
Fraction number
8) 82 84 a; 88 90 92
MkDa
43
29
18
14
8146
3
b
Fig. 20.3. (a) Chromatographic profile of TthS14 purification on a CM-Sepharose column (4 cm x 10
cm). Flow rate 70 ml/h. Fraction volume 4 ml. The total protein amount applied was 800 mg. (-0-)
absorbance at 280nm, (-e-) concentration of NaCI in the gradient. The arrow indicates the elution
peak of TthS14. (b) SDS-PAGE (15 %-discontinuous system) of CM-Sepharose fractions 80-92 ( see
Fig. 20.2. containing the TthS 14 protein. 50 ~l of each fraction were loaded on the gel. M: molecular
weight markers