BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3481

Peptides:

Sequences	Affinity Values
ERTYSPSTAVRS PSTAVR SPSTAVRS TYSPSTAVRSAA PSTETR LPSTETRHV YSPSTETRSA	[1.458 ± 0.28] [NA] [NA] [NA] [NA] [NA] [NA]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 31132884

Target Name:

The catalytic domain of rho-associated protein kinase 1

Template Name:

Not determined.

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:

Affinity Measurement Description:

The strength of interactions between Peptide7 and ROCK1 and ROCK2 was then alyzed by performing an SPR study. The binding of Peptide7 to ROCK1 as compared to ROCK2 demonstrated small differences even though bith proteins have 92% similarity in sequence in the catalytic domain. Upon inspection, there are only 3 amino acid differences in the activation loop. The average of three experiments yielded KD values of 1.458 ± 0.28 µM for ROCK1 (1–553) and 5.153 ± 1.15 µM for ROCK2.

Brief Description:

Phage particles were first mixed with MBP protein to avoid the negative selection by elimiting phages that could bind to MBP. The supertant which contained only non-MBP-binding phages was then incubated with MBP-ROCK1 (5–348).

BiopanningDataSet Comments:

Peptide7(ERTYSPSTAVRS), a promising ROCK inhibitory peptide for both ROCK isoforms, measured at 1.45 ± 0.28 µM for ROCK1 (1–553) and 5.15 ± 1.15 µM for ROCK2. Peptide7 reduced cellular migration in wound healing assays. The binding epitope on ROCK1 was mapped to the flexible activation loop within the catalytic domain. Peptide alanine scanning mutants helped identify critical amino acids to generate optimized Peptide22. This compact ROCK inhibitor facilitated vascular relaxation, blocked neovascularization of endothelial cells, and inhibited MLC phosphatase phosphorylation.

Curator:

Chunchao Pu

Curation Date:

2020-12-01

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