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Articles in PresS, published online ahead of print December 5, 2001
Am J Physiol Cell Physiol, 10.1152/ajpcell.00349.2001
Submitted on July 26, 2001
Accepted on December 4, 2001
1 Physiology, University of Kentucky, Lexington, KY, USA
* To whom correspondence should be addressed. E-mail: ejsmart{at}uky.edu.
EGF and PDGF receptors have been reported to signal via caveolin-containing membranes called caveolae, in contrast others report that EGF and PDGF receptors are exclusively associated with caveolin-devoid membranes called rafts. Our subcellular fractionation and co-immunoprecipitation studies demonstrate that in the absence of ligand, EGF and PDGF receptors are associated with rafts. However, in the presence of ligand, EGF and PDGF receptors transiently associate with caveolae. Surprisingly, pretreatment of cells with EGF prevents PDGF-dependent phosphorylation of PDGF receptors and ERK1/2 kinase activation. Furthermore, cells pretreated with PDGF prevent EGF-dependent phosphorylation of EGF receptors and ERK1/2 kinase activation. Radioligand binding studies demonstrate that incubation of cells with EGF or PDGF cause both EGF and PDGF receptors to be reversibly sequestered from the extracellular space. Experiments with methyl-ß-cyclodextrin, filipin, and anti-sense caveolin-1 demonstrate that sequestration of the receptors is dependent on cholesterol and caveolin-1. We conclude that ligand-induced stimulation of EGF or PDGF receptors can cause the heterologous desensitization of the other receptor by sequestration in cholesterol rich, caveolin-containing membranes or caveolae.
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