The interaction of an epidermal growth factor/transforming growth factor α tail chimera with the human epidermal growth factor receptor reveals unexpected complexities

Puddicombe, Sarah M., Wood, Lynn, Chamberlin, Stephen G. and Davies, Donna E. (1996) The interaction of an epidermal growth factor/transforming growth factor α tail chimera with the human epidermal growth factor receptor reveals unexpected complexities. Journal of Biological Chemistry, 271 (48), 30392-30397. (doi:10.1074/jbc.271.48.30392).

Abstract

It has been assumed that substitution of homologous regions of transforming growth factor α (TGF-α) into epidermal growth factor (EGF) can be used to probe ligand-receptor recognition without detrimental effects on ligand characteristics for the human EGF receptor (EGFR). We show that a chimera of murine (m) EGF in which the carboxyl-terminal tail is substituted for that of TGF-α (mEGF/TGF-α_44-50) results in complex features that belie this initial simplistic assumption. Comparison of EGF and mEGF/TGF- α_44-50 in equilibrium binding assays showed that although the relative binding affinity of the chimera was reduced 80-200-fold, it was more potent than EGF in mitogenesis assays using NR6/HER cells. This superagonist activity could not be attributed to differences in ligand processing or to binding to other members of the c-erbB family. It appeared to be due, in part, to choice of an EGFR-overexpressing target cell where high receptor number compensated for the low affinity of the ligand; it also appeared to be related to the ability of the chimera to activate the EGFR tyrosine kinase. Thus, when EGFR autophosphorylation was measured, mEGF/TGF-α_44-50 was more potent than EGF, despite its low affinity. When tested using chicken embryo fibroblasts, substitution of the TGF-α carboxyl-terminal tail into mEGF failed to enhance its binding affinity for chicken EGFRs; however, the chimera was intermediate in potency between TGF-α and mEGF in mitogenesis assays. Our results suggest a contextual requirement for EGFR recognition which is ligand-specific. Further, the unpredictable responses to chimeric ligands underline the complex nature of the processes of ligand recognition, receptor activation, and the ensuing cellular response.

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Contributors

Author: Donna E. Davies

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