CR1/CR2 interactions modulate the functions of the cell surface epidermal growth factor receptor

Francesca Walker, Suzanne G. Orchard, Robert Jorissen, Nathan E. Hall, Hui Hua Zhang, Peter A. Hoyne, Timothy E. Adams, Terrance G. Johns, Colin Ward, Thomas P.J. Garrett, Hong Jian Zhu, Maureen Nerrie, Andrew M. Scottt, Edouard C. Nice, Antony W. Burgess

Research output: Contribution to journalArticle

69 Citations (Scopus)

Abstract

Recent crystallographic data on the isolated extracellular domain of the epidermal growth factor receptor (EGFR) have suggested a model for its activation by ligand. We have tested this model in the context of the full-length EGFR displayed at the cell surface, by introducing mutations in two regions (CR1 and CR2) of the extracellular domain thought to be critical for regulation of receptor activation. Mutations in the CR1 and CR2 domains have opposing effects on ligand binding affinity, receptor dimerization, tyrosine kinase activation, and signaling competence. Tyr246 is a critical residue in the CR1 loop, which is implicated in the positioning and stabilization of the receptor dimer interface after ligand binding; mutations of Tyr246 impair or abolish receptor function. Mutations in CR2, which weaken the interaction that restricts the receptor to the tethered (inactive) state, enhance responsiveness to EGF by increasing affinity for the ligand. However, weakening of the CR1/CR2 interaction does not result in spontaneous activation of the receptors' kinase. We have used an antibody (mAb 806), which recognizes a transition state of the EGF receptor between the negatively constrained, tethered state and the fully active back-to-back dimer conformation, to follow conformational changes in the wild-type and mutant EGF receptors after ligand binding. Our results suggest that EGFR on the cell surface can be untethered, but this form is inactive; thus, untethering of the receptor is not sufficient for activation, and ligand binding is essential for the correct positioning of the two receptor subunits to achieve kinase activation.

LanguageEnglish
Pages22387-22398
Number of pages12
JournalJournal of Biological Chemistry
Volume279
Issue number21
DOIs
Publication statusPublished - 21 May 2004
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Walker, F., Orchard, S. G., Jorissen, R., Hall, N. E., Zhang, H. H., Hoyne, P. A., ... Burgess, A. W. (2004). CR1/CR2 interactions modulate the functions of the cell surface epidermal growth factor receptor. Journal of Biological Chemistry, 279(21), 22387-22398. https://doi.org/10.1074/jbc.M401244200
Walker, Francesca ; Orchard, Suzanne G. ; Jorissen, Robert ; Hall, Nathan E. ; Zhang, Hui Hua ; Hoyne, Peter A. ; Adams, Timothy E. ; Johns, Terrance G. ; Ward, Colin ; Garrett, Thomas P.J. ; Zhu, Hong Jian ; Nerrie, Maureen ; Scottt, Andrew M. ; Nice, Edouard C. ; Burgess, Antony W. / CR1/CR2 interactions modulate the functions of the cell surface epidermal growth factor receptor. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 21. pp. 22387-22398.
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abstract = "Recent crystallographic data on the isolated extracellular domain of the epidermal growth factor receptor (EGFR) have suggested a model for its activation by ligand. We have tested this model in the context of the full-length EGFR displayed at the cell surface, by introducing mutations in two regions (CR1 and CR2) of the extracellular domain thought to be critical for regulation of receptor activation. Mutations in the CR1 and CR2 domains have opposing effects on ligand binding affinity, receptor dimerization, tyrosine kinase activation, and signaling competence. Tyr246 is a critical residue in the CR1 loop, which is implicated in the positioning and stabilization of the receptor dimer interface after ligand binding; mutations of Tyr246 impair or abolish receptor function. Mutations in CR2, which weaken the interaction that restricts the receptor to the tethered (inactive) state, enhance responsiveness to EGF by increasing affinity for the ligand. However, weakening of the CR1/CR2 interaction does not result in spontaneous activation of the receptors' kinase. We have used an antibody (mAb 806), which recognizes a transition state of the EGF receptor between the negatively constrained, tethered state and the fully active back-to-back dimer conformation, to follow conformational changes in the wild-type and mutant EGF receptors after ligand binding. Our results suggest that EGFR on the cell surface can be untethered, but this form is inactive; thus, untethering of the receptor is not sufficient for activation, and ligand binding is essential for the correct positioning of the two receptor subunits to achieve kinase activation.",
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Walker, F, Orchard, SG, Jorissen, R, Hall, NE, Zhang, HH, Hoyne, PA, Adams, TE, Johns, TG, Ward, C, Garrett, TPJ, Zhu, HJ, Nerrie, M, Scottt, AM, Nice, EC & Burgess, AW 2004, 'CR1/CR2 interactions modulate the functions of the cell surface epidermal growth factor receptor', Journal of Biological Chemistry, vol. 279, no. 21, pp. 22387-22398. https://doi.org/10.1074/jbc.M401244200

CR1/CR2 interactions modulate the functions of the cell surface epidermal growth factor receptor. / Walker, Francesca; Orchard, Suzanne G.; Jorissen, Robert; Hall, Nathan E.; Zhang, Hui Hua; Hoyne, Peter A.; Adams, Timothy E.; Johns, Terrance G.; Ward, Colin; Garrett, Thomas P.J.; Zhu, Hong Jian; Nerrie, Maureen; Scottt, Andrew M.; Nice, Edouard C.; Burgess, Antony W.

In: Journal of Biological Chemistry, Vol. 279, No. 21, 21.05.2004, p. 22387-22398.

Research output: Contribution to journalArticle

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T1 - CR1/CR2 interactions modulate the functions of the cell surface epidermal growth factor receptor

AU - Walker, Francesca

AU - Orchard, Suzanne G.

AU - Jorissen, Robert

AU - Hall, Nathan E.

AU - Zhang, Hui Hua

AU - Hoyne, Peter A.

AU - Adams, Timothy E.

AU - Johns, Terrance G.

AU - Ward, Colin

AU - Garrett, Thomas P.J.

AU - Zhu, Hong Jian

AU - Nerrie, Maureen

AU - Scottt, Andrew M.

AU - Nice, Edouard C.

AU - Burgess, Antony W.

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