BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 2485

Peptides:

Sequences
CWDDGWLC(131) CWDDLWWLC(10) CWDDGLMC(7) CWDDGWMC(4)

Sequences Features:

Motif:

CWDD[GL]WLC

Unique Sequence Number:

Experimental Method:

Phage display (competitive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 7657703

Target Name:

Fibronectin type Ⅲ10(RGD)-containing fragments

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

CX5C+CX6C+CX7C+CX9 phage display library pool

Affinity Measurement Method:

Affinity Measurement Description:

Brief Description:

After the fourth panning, wells with bound phages were eluted with 1 mM solution of GRGDSP or *CELRGDGWC* peptidee.

BiopanningDataSet Comments:

A predominant cyclic motif, *CWDD[GL]WLC*, was obtained (the asterisks denote a potential disulfide bond). Studies using the purified phage and the corresponding synthetic cyclic peptides showed that *CWDDGWLC*-expressing phage binds specifically to fibronectin and to fibronectin fragments containing the RGD sequence. The binding did not require divalent cations and was inhibited by both RGD and *CWDDGWLC*-containing synthetic peptides. Conversely, RGD-expressing phage attached specifically to immobilized *CWDDGWLC*-peptide and the binding could be blocked by the respective synthetic peptides in solution. Moreover, fibronectin bound to a *CWDDGWLC*-peptide affinity column, and could be eluted with an RGD-containing peptide. The *CWDDGWLC*-peptide inhibited RGD-dependent cell attachment to fibronectin and vitronectin, but not to collagen.A region of the β3 subunit of RGD-binding integrins that has been previously demonstrated to be involved in ligand binding includes a polypeptide stretch, KDDLW (in β3) similar to WDDC/LWL. Synthetic peptides corresponding to this region in β3 were found to bind RGD-displaying phage and conversion of its two aspartic residues into alanines greatly reduced the RGD binding. Polyclonal antibodies raised against the *CWDDGWLC*-peptide recognized β1 and β3 in immunoblots. These data indicate that the *CWDDGWLC*-peptide is a functional mimic of ligand binding sites of RGD-directed integrins, and that the structurally similar site in the integrin β subunit is a binding site for RGD.

Curator:

Curation Date:

0000-00-00

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