scholarly journals The membrane proximal disulfides of the EGF receptor extracellular domain are required for high affinity binding and signal transduction but do not play a role in the localization of the receptor to lipid rafts

2006 ◽  
Vol 1763 (8) ◽  
pp. 870-878 ◽  
Author(s):  
Jennifer Macdonald ◽  
Zhengzhe Li ◽  
Wanwen Su ◽  
Linda J. Pike
Keyword(s):  
Signal Transduction ◽  
Lipid Rafts ◽  
Egf Receptor ◽  
Extracellular Domain ◽  
Affinity Binding ◽  
High Affinity Binding ◽  
High Affinity

scholarly journals Cloning of a murine IL-11 receptor alpha-chain; requirement for gp130 for high affinity binding and signal transduction.

1994 ◽  
Vol 13 (20) ◽  
pp. 4765-4775 ◽  
Author(s):  
D.J. Hilton ◽  
A.A. Hilton ◽  
A. Raicevic ◽  
S. Rakar ◽  
M. Harrison-Smith ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Affinity Binding ◽  
High Affinity Binding ◽  
Receptor Alpha ◽  
Alpha Chain ◽  
High Affinity ◽  
Receptor Alpha Chain

scholarly journals Kinetics of binding, endocytosis, and recycling of EGF receptor mutants

1992 ◽  
Vol 117 (1) ◽  
pp. 203-212 ◽  
Author(s):  
S Felder ◽  
J LaVin ◽  
A Ullrich ◽  
J Schlessinger
Keyword(s):  
Point Mutation ◽  
Phorbol Ester ◽  
Egf Receptor ◽  
Receptor Internalization ◽  
Affinity Binding ◽  
High Affinity Binding ◽  
High Affinity ◽  
Low Concentrations ◽  
Internalization Rate ◽  
Kinetics Of

This report describes analysis of factors which regulate the binding of EGF to EGF receptor, receptor internalization, and receptor recycling. Three different methods were used to inhibit high-affinity EGF binding as measured at equilibrium: treatment of cells with an active phorbol ester (PMA), binding of a mAb directed against the EGF receptor (mAb108), and truncation of most of the cytoplasmic domain of the receptor. These treatments reduced the rate at which low concentrations of EGF bound to cells, but did not affect the rate of EGF dissociation. We conclude that high-affinity EGF binding on living cells results from a difference in the apparent on rate of EGF binding. We then used these conditions and cell lines to test for the rate of EGF internalization at different concentrations of EGF. We demonstrate that internalization of the EGF receptor is stimulated roughly 50-fold at saturating concentrations of EGF, but is stimulated an additional two- to threefold at low concentrations (less than 1 nM). Four treatments reduce the rate of internalization of low concentrations of EGF to the rate seen at saturating EGF concentrations. Phorbol ester treatment and mAb108 binding to "wild type" receptor reduce this rate (and reduce high-affinity binding). Point mutation at Lys721 (kinase negative EGF receptor) and point mutation at Thr654 (removing a major site of protein kinase C phosphorylation) reduce the internalization rate, without affecting high-affinity binding. We suggest that while EGF stimulates endocytosis for all receptors, high-affinity receptors bind and are internalized more quickly than low-affinity receptors. Tyrosine kinase activity and the Thr654 region appear necessary for this response.

The extracellular domain of the neurokinin-1 receptor is required for high-affinity binding of peptides

Biochemistry ◽  
1992 ◽  
Vol 31 (47) ◽  
pp. 11806-11811 ◽  
Author(s):  
Tung Ming Fong ◽  
Hong Yu ◽  
Ruey Ruey C. Huang ◽  
Catherine D. Strader
Keyword(s):  
Extracellular Domain ◽  
Affinity Binding ◽  
High Affinity Binding ◽  
Neurokinin 1 Receptor ◽  
High Affinity ◽  
Neurokinin 1

Heterodimerization of the epidermal-growth-factor (EGF) receptor and ErbB2 and the affinity of EGF binding are regulated by different mechanisms

2001 ◽  
Vol 356 (1) ◽  
pp. 87-96 ◽  
Author(s):  
Lene E. JOHANNESSEN ◽  
Karianne E. HAUGEN ◽  
Anne Carine ØSTVOLD ◽  
Espen STANG ◽  
Inger Helene MADSHUS
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Egf Receptor ◽  
Kinase Inhibitor ◽  
Affinity Binding ◽  
High Affinity Binding ◽  
Receptor Dimerization ◽  
High Affinity ◽  
Epidermal Growth ◽  
Specific Kinase Inhibitor

When clathrin-dependent endocytosis is inhibited in HeLa cells by overexpression of a K44A (Lys44 → Ala) mutant of the GTPase dynamin, high-affinity binding of epidermal growth factor (EGF) to the EGF receptor (EGFR) is disrupted [Ringerike, Stang, Johannessen, Sandnes, Levy and Madshus (1998) J. Biol. Chem. 273, 16639–16642]. We now report that the effect of [K44A]dynamin on EGF binding was counteracted by incubation with the non-specific kinase inhibitor staurosporine (SSP), implying that a protein kinase is responsible for disrupted high-affinity binding of EGF upon overexpression of [K44A]dynamin. The effect of [K44A]dynamin on EGF binding was not due to altered phosphorylation of the EGFR, suggesting that the activated kinase is responsible for phosphorylation of a substrate other than EGFR. The number of EGFR molecules was increased in cells overexpressing [K44A]dynamin, while the number of proto-oncoprotein ErbB2 molecules was unaltered. EGF-induced receptor dimerization was not influenced by overexpression of [K44A]dynamin. ErbB2–EGFR heterodimer formation was found to be ligand-independent, and the number of heterodimers was not altered by overexpression of [K44A]dynamin. Neither SSP nor the phorbol ester PMA, which disrupts high-affinity EGF–EGFR interaction, had any effect on the EGFR homo- or hetero-dimerization. Furthermore, the EGF-induced tyrosine phosphorylation of ErbB2 was not affected by overexpression of [K44A]dynamin, implying that EGFR–ErbB2 dimers were fully functional. Our results strongly suggest that high-affinity binding of EGF and EGFR–ErbB2 heterodimerization are regulated by different mechanisms.

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