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Argos inhibits epidermal growth factor receptor signalling by ligand sequestration

Abstract

The epidermal growth factor receptor (EGFR) has critical functions in development and in many human cancers1,2,3. During development, the spatial extent of EGFR signalling is regulated by feedback loops comprising both well-understood activators and less well-characterized inhibitors3,4. In Drosophila melanogaster the secreted protein Argos functions as the only known extracellular inhibitor of EGFR5, with clearly identified roles in multiple stages of development3. Argos is only expressed when the Drosophila EGFR (DER) is activated at high levels6, and downregulates further DER signalling. Although there is ample genetic evidence that Argos inhibits DER activation, the biochemical mechanism has not been established. Here we show that Argos inhibits DER signalling without interacting directly with the receptor, but instead by sequestering the DER-activating ligand Spitz. Argos binds tightly to the EGF motif of Spitz and forms a 1:1 (Spitz:Argos) complex that does not bind DER in vitro or at the cell surface. Our results provide an insight into the mechanism of Argos function, and suggest new strategies for EGFR inhibitor design.

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Figure 1: Quantitative analysis of Spitz/Argos interactions.
Figure 2: Argos sequesters Spitz away from DER.
Figure 3: Argos does not displace Spitz pre-bound to cell surface DER.

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Acknowledgements

We thank J. Duffy, K. Ferguson, P. Carroll, G. Van Duyne and members of the Lemmon and Shvartsman laboratories for valuable discussions; T. Schüpbach and N. Perrimon for providing cDNAs. This work was supported by grants from the NIH (to M.A.L.) and NSF (S.Y.S.), by NIH training grant support (to D.E.K. and V.M.N.), and by an NSF graduate research fellowship (G.T.R.).

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Correspondence to Mark A. Lemmon.

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Supplementary information

Supplementary Figure 1

SPR binding curves showing that the Argos/Spitz interaction requires only the EGF motif of Spitz and the C-terminal 225 amino acids of Argos. (DOC 365 kb)

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Klein, D., Nappi, V., Reeves, G. et al. Argos inhibits epidermal growth factor receptor signalling by ligand sequestration. Nature 430, 1040–1044 (2004). https://doi.org/10.1038/nature02840

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