| Title | Induction of notch signaling by immobilization of jagged-1 on self-assembled monolayers |
| Publication Type | Journal Article |
| 2009 |
| Authors | Gonçalves, RM, Martins, MCL, Almeida-Porada, G, Barbosa, MA |
| Journal | BiomaterialsBiomaterials |
| Volume | 30 |
| Issue | 36 |
| Pagination | 6879 - 6887 |
| Date Published | 2009/// |
| 01429612 (ISSN) |
| Activator concentration, Adequate models, Adsorption, Animals, article, Bioactivity, Biological activities, Biological materials, biomaterial, Calcium-Binding Proteins, Carbonyldiimidazole, Cell culture, Cell Line, Cell membranes, cell strain HL 60, Cell system, Chemical activation, Concentration (process), Concentration of, controlled study, Covalently bound, Ethylene, Ethylene Glycol, gene expression, gold, Gold substrates, HL-60 cell line, human, human cell, Human IgG, Humans, Immunoassay, immunoglobulin Fc fragment, Immunoglobulin G, immunoglobulin G antibody, Intercellular Signaling Peptides and Proteins, Jagged1, Ligands, Mammalia, Membrane Proteins, Model surface, Molecular Structure, Nanostructured materials, Nanostructured surfaces, Nonfouling, Notch receptor, Notch signaling, Photoelectron spectroscopy, priority journal, Protein adsorption, protein immobilization, Radiolabeling, Real-time PCR, Receptors, Fc, Receptors, Notch, Recombinant Fusion Proteins, Sams, self assembled monolayer, Self assembled monolayers, Self-assembled monolayers, signal transduction, Signaling, stem cell, Surface Properties, Target genes, Tetra, transcription factor HES 1, X ray photoelectron spectroscopy |
| 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. © 2009 Elsevier Ltd. |
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