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Induction of notch signaling by immobilization of jagged-1 on self-assembled monolayers

Paper ID Volume ID Publish Year Pages File Format Full-Text
8770 605 2009 9 PDF Available
Title
Induction of notch signaling by immobilization of jagged-1 on self-assembled monolayers
Abstract

Notch signaling is a key mechanism during mammal development and stem cell regulation. This study aims to target and control Notch signaling by ligands immobilization using self-assembled monolayers (SAMs) as model surfaces. Non-fouling substrates were prepared by immersion of gold substrates in (1-Mercapto-11-undecyl)tetra(ethylene glycol) thiol solutions. These surfaces were activated with N,N′-carbonyldiimidazole (CDI) at different concentrations (0, 0.03, 0.3, 3 and 30 mg/ml) and an anti-human IgG, Fc specific fragment antibody (Ab) was covalently bound to EG4-SAMs to guarantee the correct exposure of the Notch ligand Jagged-1/Fc chimera (Jag-1). The presence of Ab and Jag-1 was confirmed by radiolabeling, X-ray photoelectron spectroscopy (XPS), ellipsometry and ELISA. The biological activity of Jag-1-Ab-SAMs was assessed by real-time PCR for Hes-1 family gene expression, a Notch pathway target gene, in HL-60 cell line. Results have shown an increase of the amount of immobilized Ab with increasing surface activator concentrations. Jag-1 concentration also increases with Ab concentration. Interestingly, a higher Jagged-1 exposure and fold increase in Hes-1 expression were obtained for surfaces activated with the lowest concentration of CDI (0.03 mg/ml). These results illustrate the great importance of ligands orientation and exposure, when compared with density. This investigation brings new insights into Notch signaling mechanisms. In particular, Jag-1-Ab-SAMs have shown to be adequate model surfaces to study Notch pathway activation and may provide a basis to develop new interfaces in biomaterials to control Notch mechanism in different cell systems.

Keywords
Nanostructured surfaces; Self-assembled monolayers; Protein immobilization; Protein adsorption; Notch signaling
First Page Preview
Induction of notch signaling by immobilization of jagged-1 on self-assembled monolayers
Publisher
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 30, Issue 36, December 2009, Pages 6879–6887
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering