BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3000

Peptides:

Sequences

CKMTRSTIC(29)
CKIMISISC(19)
CLIRRTSIC(8)
CPLLTKLKC(8)
CHIPSIHSC(8)
CTHRRSTPC(7)
CLHPPLTLC(6)
CNLLLRTQC(5)
CQMMLIRRC(4)
CPRRSHPIC(4)
CILRKIRPC(4)
CKMNIRLSC(2)
CSRRPTSIC(1)
CLIRLILQC(1)
CRMRKTLRC(1)
CQPHHSIIC(1)
CSTHIPSHC(1)
CILRRRKRC(1)
CSIRIHRRC(1)
CSLRRHPIC(1)
CRTKLRKLC(1)
CHMQIMHRC(1)
CTRMMLRIC(1)
CTPRQTRMC(1)
CMLNITRRC(1)
CRKLPRISC(1)
CPLQPKSIC(1)
CPIRHRPIC(1)
CRTMRIRLC(1)

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (in vivo)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 27173134

Target Name:

Endothelial cells (ECs) of ligated left carotid artery (LCA)

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

Ph.D.-C7C phage display library (CX7C)

Affinity Measurement Method:

Affinity Measurement Description:

Brief Description:

Briefly, 1.0e11 plaque-forming units (pfu) of phages were injected intravenously into mice at 3 days after left carotid artery (LCA) ligation. After 10 minutes of circulation, the mice were anesthetized with zoletil and rompun, and pressure perfused with saline containing heparin (10 U/ml) via the left ventricle after severing the inferior vena cava to remove unbound phage. The LCA was then isolated. The carotid lumen was rapidly flushed using a 29-gauge insulin syringe with 200 μL of phage elution buffer (0.2M Glycine-HCl, pH 2.2) for detachment of bound phages from endothelium and subsequently neutralized with 30 μL of 1 M Tris-HCl (pH 9.1) using a 29-gauge insulin syringe into a microfuge tube. Eluted phages from LCA were amplified in Escherichia coli ER2738 for reinjection in the next round.

BiopanningDataSet Comments:

Two peptides, CLIRRTSIC and CPRRSHPIC, selectively bound to arterial endothelial cells (ECs) exposed to disturbed flow not only in the partially ligated carotids but also in the lesser curvature and branching point of the aortic arch in mice as well as human pulmonary artery branches. Peptides were conjugated to branched polyethyleniminepolyethylene glycol polymer to generate polyplexes carrying siRNA targeting intercellular adhesion molecule-1 (siICAM-1). In mouse model, CLIRRTSIC polyplexes carrying si-ICAM-1 specifically bound to endothelium in disturbed flow regions, reducing endothelial ICAM-1 expression. Mass spectrometry analysis revealed that non-muscle myosin heavy chain II A (NMHC IIA) is a protein targeted by CLIRRTSIC peptide. Further studies showed that shear stress regulates NMHC IIA expression and localization in ECs. The CLIRRTSIC is a novel peptide that could be used for targeted delivery of therapeutics such as siRNAs to pro-atherogenic endothelium.

Curator:

Curation Date:

0000-00-00

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