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.