Superagonistic Activation of ErbB-1 by EGF-related Growth Factors with Enhanced Association and Dissociation Rate Constants

Epidermal growth factor (EGF) and transforming growth factor-α (TGFα) are mitogenic hormones that exert their activity primarily by binding to the EGF receptor, also known as ErbB-1. We have recently characterized a set of EGF/TGFα chimeric molecules with similar high affinity for ErbB-1 as EGF and TGFα and shown that three of these chimeras induce mitogenic cell stimulation at already a 10-fold lower concentration than their wild-type counterparts (Lenferink, A. E., Kramer, R. H., van Vugt, M. J., Königswieser, M., DiFiore, P. P., van Zoelen, E. J., and van de Poll, M. L. (1997) Biochem. J. 327, 859–865). In the present study we show that these so-called superagonistic chimeras do not differ from EGF and TGFα in their ability to induce ErbB-1 tyrosine phosphorylation but are considerably more potent in activation of mitogen-activated protein kinase phosphorylation. Direct cell binding studies and analysis of ligand-receptor interaction by surface plasmon resonance measurements revealed that both the association rate constant ( k on ) and the dissociation rate constant ( k off ) of these superagonists is 3–5-fold higher in comparison with the wild-type ligands and nonsuperagonistic chimeras. These data indicate that the dynamic on and off rate constants for receptor binding may be more specific parameters for determining the mitogenic activity of peptide hormones than their constants for equilibrium receptor binding.