Identification of the Minimal Requirements for Binding to the Human Epidermal Growth Factor (EGF) Receptor Using Chimeras of Human EGF and an EGF Repeat of Drosophila Notch

Many proteins contain so-called epidermal growth factor (EGF)-like domains that share the characteristic spacing of cysteines and glycines with members of the EGF family. They are, however, functionally unrelated, despite the fact that the three-dimensional structure of these EGF-like domains, also,...

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Bibliographic Details
Published in:The Journal of biological chemistry 1998-06, Vol.273 (26), p.16075-16081
Main Authors: van de Poll, M L, van Vugt, M J, Lenferink, A E, van Zoelen, E J
Format: Article
Language:English
Subjects:
Animals
Cells, Cultured
Drosophila
Drosophila Proteins
Epidermal Growth Factor - genetics
Epidermal Growth Factor - metabolism
ErbB Receptors - metabolism
Humans
Ligands
Magnetic Resonance Spectroscopy
Membrane Proteins - genetics
Membrane Proteins - metabolism
Protein Structure, Secondary
Receptors, Cell Surface - genetics
Receptors, Notch
Recombinant Fusion Proteins - metabolism
Structure-Activity Relationship
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Summary:Many proteins contain so-called epidermal growth factor (EGF)-like domains that share the characteristic spacing of cysteines and glycines with members of the EGF family. They are, however, functionally unrelated, despite the fact that the three-dimensional structure of these EGF-like domains, also, is often very similar to that of the EGF receptor agonists. In the present study, we linked an EGF-like repeat from the Drosophila Notch protein to the N- and C-terminal linear tail sequences of human EGF (hEGF), and we showed that this chimera (E1N6E) is unable to bind or activate the hEGF receptor. This recombinant protein was then used as a basic construct for identifying the minimal requirements for high affinity EGF receptor binding and activation. We selectively reintroduced a limited number of important hEGF-derived residues, and by using this unique approach, we were able to make hEGF/Notch chimeras that, compared with wild type hEGF, showed nearly 100% binding affinity and mitogenic activity on HER-14 cells expressing the hEGF receptor.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.273.26.16075