The Folding Pathway of Disulfide Containing Proteins
195
GSSG (0.5 mM)
Cys-Cys (O.5 mM)
120
120
R+I+II
III ..........
N
N
60
60
GSSG (2 mM)
Cys-Cys (2 mM)
120
120
R+I+II
R+I+II
III-..
III
N
N
60
60
GSH / GSSG (4 mM / 2 mM)
Cys / Cys-Cys (4 mM / 2 mM)
~
120~----------------------'
?f. '-"
R+I+II
120~---------------------'
R+I+II
N
N
III
III
•
15
60
180
2 5
15
60
180
Time of folding (min)
Fig. 13.3. Effects of redox agents on the kinetics of flow of the folding intermediates of PCI.. Foldings
were performed in the Tris-HCl buffer containing the indicated redox agents. I, II and III stand for 1disulfide, 2-disulfide and 3-disulfide scrambled species of PCI, respectively. [R+I+II], [III] and [N]
were followed quantitatively in each time-course trapped sample analyzed by HPLC. [R], [I] and [II]
overlap extensively, therefore, their recoveries were quantitated collectively