Epidermal growth factor tethered through coiled-coil interactions induces cell surface receptor phosphorylation.
We have elaborated and validated a novel approach for the oriented tethering of proteins such as the epidermal growth factor (EGF) on aminated surfaces. The grafting reactions were optimized to generate a dense and homogeneous EGF layer. Impact of EGF orientation on A-431 cellular response was investigated. Our results demonstrate that, in sharp contrast to responses obtained with soluble EGF supply or with randomly grafted EGF, oriented immobilization of EGF via a de novo designed coiled-coil capture system leads to a sustained phosphorylation of A-431 cell surface EGF receptors. Our results thus indicate that oriented protein immobilization via coiled-coil interactions is an efficient and versatile method to control tethering of bioactive molecules for future applications in the field of regenerative medicine and tissue engineering.
Boucher
Cyril
C
Department of Chemical Engineering, Groupe de Recherche en Sciences et Technologie Biomédicales (GRSTB), Ecole Polytechnique de Montréal, Montréal (Qc), Canada.
Liberelle
Benoît
B
Jolicoeur
Mario
M
Durocher
Yves
Y
De Crescenzo
Gregory
G
D002478:Cells, Cultured; D004815:Epidermal Growth Factor / Q000378:metabolism*; D006801:Humans; D015394:Molecular Structure; D010766:Phosphorylation; D011437:Propylamines; D011956:Receptors, Cell Surface / Q000378:metabolism*; D012821:Silanes / Q000737:chemistry; D012825:Silicon / Q000737:chemistry; D013499:Surface Properties
D011437:Propylamines; D011956:Receptors, Cell Surface; D012821:Silanes; D004815:Epidermal Growth Factor; C477625:amino-propyl-triethoxysilane; D012825:Silicon