Biomedical Engineering Reference
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(a)
Before blocking
18
16
14
12
10
8
6
4
2
0
AZ
AZ
After blocking
p53
p53
p53
Au
(b)
Before blocking
15
After blocking
AZ
Mdm2
10
p53
5
p53
p53
Au
0
(c)
Before blocking
After blocking
15
Mdm2
AZ
10
5
p53
p53
p53
Au
0
FIGURE 6.8
(See color insert.)
Competitive blocking experiments on the p53-mdm2-
azurin ternary complex. (a) Azurin is used as a competitor for the p53-azurin complex (left);
unbinding frequencies before and after blocking the substrate with a solution of free azurin
(right). (b) mdm2 is used as a competitor for the p53-azurin complex (left); unbinding frequen-
cies before and after blocking the substrate with a solution of free mdm2 (right). (c) Azurin
is used as a competitor for the p53-mdm2 complex (left); unbinding frequencies before and
after blocking the substrate with a solution of free azurin (right). (Adapted from Bizzarri, A.
R. and Cannistraro, S. 2009.
J. Phys. Chem. B
, 113, 16449-16464. Adapted from Funari, G.
et al. 2010.
J. Mol. Recognit.
, 23, 343-351).
carried out by measuring the frequency of the unbinding events between p53 immo-
bilized on the substrate and mdm2 anchored to the tip before and after blocking
the substrate with a solution of free azurin. In both the latter cases, substantially no
changes in the unbinding frequency has been observed. Such a finding has provided
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