Epidermal growth factor mutant with wild-type affinity for both ErbB1 and ErbB3.
The family of epidermal growth factor (EGF)-like ligands binds to ErbB receptors in a highly selective manner. Previous studies indicated that both linear regions of the ligand play a major role in determining receptor selectivity, and phage display studies showed that each region could be optimized independently for enhanced affinity. In this study, we broadened the ErbB binding specificity of EGF by introducing the optimal sequence requirements for ErbB3 binding in both the N- and C-terminal linear regions. One such EGF mutant, designated WVR/EGF/IADIQ, gained high affinity for ErbB3 and showed concomitant ErbB3 activation through ErbB2.ErbB3 heterodimers similar to the natural ErbB3 ligand NRG1beta, while the capacity to bind and activate ErbB1 was fully maintained. Despite its high affinity for ErbB1 and ErbB3, this mutant was unable to activate ErbB1.ErbB3 heterodimers, as shown by the cell survival and receptor phosphorylation analysis. We concluded that despite the fact that no naturally occurring ligand exists with this dual-specificity, high-affinity binding to both ErbB1 and ErbB3 is not mutually exclusive. This mutant can be useful in a direct structural comparison of the ligand-binding characteristics of ErbB1 and ErbB3.
Department of Cell Biology, Radboud University Nijmegen, Faculty of Science, Toernooiveld 1, 6525 ED Nijmegen, The Netherlands.
Miriam
M
Wingens
Saskia
S
Jacobs-Oomen
Sebastian P
SP
van der Woning
Catelijne
C
Stortelers
Everardus J J
EJ
van Zoelen
D000818:Animals; D001667:Binding, Competitive; D002460:Cell Line; D019281:Dimerization; D004815:Epidermal Growth Factor / Q000235:genetics* / Q000378:metabolism*; D066246:ErbB Receptors / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D051379:Mice; D020836:Protein Structure, Quaternary; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism; D020893:Receptor, ErbB-3 / Q000378:metabolism*
D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D008024:Ligands; D018719:Erb-b2 Receptor Tyrosine Kinases; D020893:Receptor, ErbB-3