An N-terminal deletion mutant of Fip1 in which this binding site is disrupted cannot complement the loss of wild-type Fip1, but the mutant is fully functional if it is fused directly to PAP.
A mouse line of mutated gp130 in which the SHP-2/SOCS3-binding site was disrupted developed a RA-like joint disease with increased production of Th1-type cytokines and immunoglobulins of the IgG2a and IgG2b classes [ 78].
When both Egr-1 binding sites were disrupted as in pRad-155m3, the basal level of the Rad promoter activity dropped dramatically and no induction by PDGF was found (Figure 2C).
To test this hypothesis, we constructed two mKv1.3 mutants in which the verapamil binding sites are disrupted [ 44].
We utilised our fused PCNA heterotrimer to generate mutant rings in which one of the three enzyme binding sites is disrupted at a time.
Consistent with this view, the crystal structure of the eIF4AIII-MLN51 complex in the absence of ATP, showed that eIF4AIII adopts an open conformation in which the RNA-binding site is disrupted [17].
These mutants interact as well as unmutated enzyme with the affinity column, suggesting that the binding site was not disrupted; furthermore, fluorescence scans and CD spectroscopy revealed no apparent changes in structure.
In addition, the p53 peptide containing this binding site was able to disrupt the binding of FAK and p53, to activate p53 and to inhibit viability of HCT116p53+/+ cells compared to HCT116p53-/ cells, suggesting that FAK-p53 targeting can be used for therapeutics [ 20].
When sorted by their MACS binding score, a clear trend emerged that weaker sites were disrupted at lower dose and experienced greater changes.
The ratio of P-values of affinity scores between two alleles can then be used to determine if the TF binding site is likely to be disrupted.
