BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 1653

Peptides:

Sequences
PRFWEYWLRLME(10) KSFQQYWQELML(9) KTFEEYWLMLMS(4) PSFWEHWVELML(4) KRFQDYWSELML(3)

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

mRNA display (common panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 21423613

Target Name:

E3 ubiquitin-protein ligase Mdm2

Template Name:

Cellular tumor antigen p53

Structure of Target-Template Complex:

1YCR

4HFZ

Structure of Target-Peptide Complex:

Not determined.

Library Name:

X(2)-F-X(3)-W-X(2)-L-X(2) mRNA display library

Affinity Measurement Method:

ELISA

Affinity Measurement Description:

In ELISA, MIP (PRFWEYWLRLME) could inhibit the MDM2-p53 interaction with an IC50 of 10 nM. Also MIP could inhibit the MDMX-p53 interaction at an IC50 of 120 nM.

Brief Description:

The mRNA-displayed peptide library is incubated with MDM2 immobilized on beads through an affinity selection tag containing a ZZ domain and a TEV protease cleavage site, and unbound molecules are washed away. The bound molecules are eluted by cleavage with the TEV protease, and their mRNA portion is amplified by RT-PCR. The resulting DNA can be used for the next rounds of selection or analyzed by cloning and sequencing.

BiopanningDataSet Comments:

While previous studies have used phage display to identify peptides (such as DI) that inhibit the MDM2-p53 interaction, these peptides were not sufficiently optimized because the size of the phage-displayed random peptide libraries did not cover all of the possible sequences. In this study, authors performed selection of MDM2-binding peptides from large random peptide libraries in two stages using mRNA display. Authors identified an optimal peptide (PRFWEYWLRLME) named MIP that inhibited the MDM2-p53 and MDMX-p53 interactions 29- and 13-fold more effectively than DI, respectively. Expression of MIP fused to the thioredoxin scaffold protein in living cells by adenovirus caused stabilization of p53 through its interaction with MDM2, resulting in activation of the p53 pathway. Furthermore, expression of MIP also inhibited tumor cell proliferation in a p53-dependent manner more potently than DI.

Curator:

Curation Date:

0000-00-00

First |

Prev |

1653

| Next

| Last