BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 2627

Peptides:

Sequences
WTGWCLNPEESTWGFCTGSF(9/25) LNWECWYDYRLEAWDCRGDI(4/25) DTDMCWWWSREFGWECAGAG(2/25) VVACSWDWTMGAVVCYERI(1/25)

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 15300809

Target Name:

Extracellular domain of receptor tyrosine-protein kinase erbB-2

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

X4CX10CX4 fUSE5 phage display library

Affinity Measurement Method:

Affinity Measurement Description:

Brief Description:

In the first round, 0.9 μg ECDAP were incubated with anti-AP conjugated agarose beads on a nutator overnight at 4°C. The amplified phage library was incubated with anti-AP conjugated agarose beads alone for 2 hr on a nutator at room temperature to remove those phage specific for AP. The unbound phage were filtered with a 0.45 μm cellulose acetate syringe filter and incubated with ECDAP bound to beads on a nutator for 2 hr at room temperature. The unbound phage were removed by washing 5 times with PBS containing a cocktail of prokaryotic protease inhibitors (PPI) (Sigma). Additional two rounds of screening were performed.

BiopanningDataSet Comments:

One clone (EC-1, displaying WTGWCLNPEESTWGFCTGSF) bound all preparations of ErbB2 including live cells and fresh frozen human breast cancer specimens. The synthetic peptide based on the deduced sequence of the EC-1 clone and its biotin-conjugated form retained binding affinity for purified ErbB2 and ErbB2 overexpressing cell lysates. EC-1 peptide was able to effectively inhibit the phosphorylation of ErbB2 on residues Y1248 and Y877 in a dose- and time-dependent manner. Furthermore, EC-1 peptide selectively inhibits the proliferation of ErbB2 overexpressing breast cancer cells. The linear portion of the cyclic EC-1 peptide was shown to be essential for binding ErbB2. In addition, 4 biased phage libraries were constructed allowing 4 different regions of the EC-1 peptide to have random sequence.

Curator:

Curation Date:

0000-00-00

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