BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 2438

Peptides:

Sequences	Affinity Values
SWLAYPGAVSYR(7/10) YSAYPDSVPMMS(2/10)	[0.916][678 ± 23] [0.856][186 ± 7]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (competitive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 12351647

Target Name:

Ephrin type-A receptor 2

Template Name:

Type A ephrins

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

Ph.D.-12 phage display library (X12)

Affinity Measurement Method:

ELISA,Surface plasmon resonance (SPR)

Affinity Measurement Description:

Phage ELISA was performed to measure the affinity of phage to EphA2. Absorbance at 405 nm was determined, reproduced from the graph and shown. Besides, equilibrium binding data obtained by surface plasmon resonance indicate that the YSAYPDSVPMMS peptide binds to EphA2 with higher affinity (KD = 186 nM ± 7) than the SWLAYPGAVSYR peptide (KD = 678 nM ± 23).

Brief Description:

For each round of biopanning, phages remaining bound after washing were eluted with ephrin-A1 Fc.

BiopanningDataSet Comments:

Nine of 10 phage clones from the pool eluted with ephrin-A1 bind specifically to EphA2 (data not shown). Seven of these clones display the SWLAYPGAVSYR peptide (SWL peptide) and two display the peptide YSAYPDSVPMMS (YSA peptide). The YSA peptide targets EphA2 on the cell surface and stimulates activation of the receptor. Thus, the YSA peptide is an agonist for EphA2. Besides, the YSA and SWL peptides inhibit ephrin-A binding to EphA2. To further characterize the binding site of the YSA and SWL peptides, competition experiments with phage displayed peptides were performed. Synthetic SWL peptide competes with YSA phage bound to immobilized EphA2, and conversely YSA peptide competes with SWL phage. This indicates that the YSA and SWL peptides bind to the same or overlapping sites on EphA2.

Curator:

Curation Date:

0000-00-00

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