Controlling Epidermal Growth Factor (EGF)-stimulated Ras Activation in Intact Cells by a Cell-permeable Peptide Mimicking Phosphorylated EGF Receptor
| Autor: | SongYi Yao, Yao-Zhong Lin, Mauricio Rojas |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Phosphopeptides
Cell Membrane Permeability Transcription Genetic Molecular Sequence Data Biochemistry Mice Epidermal growth factor Animals Humans Amino Acid Sequence Fluorescent Antibody Technique Indirect Protein kinase A Molecular Biology Peptide sequence Adaptor Proteins Signal Transducing GRB2 Adaptor Protein Epidermal Growth Factor biology Autophosphorylation Proteins Signal transducing adaptor protein 3T3 Cells Cell Biology Molecular biology Peptide Fragments ErbB Receptors Kinetics Calcium-Calmodulin-Dependent Protein Kinases ras Proteins biology.protein GRB2 biological phenomena cell phenomena and immunity Signal transduction hormones hormone substitutes and hormone antagonists Signal Transduction |
| Zdroj: | Journal of Biological Chemistry. 271:27456-27461 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.271.44.27456 |
| Popis: | Epidermal growth factor (EGF)-stimulated Ras activation involves specific interactions between the EGF receptor (EGFR), the adaptor proteins Grb2 and Shc, and the nucleotide exchange factor Sos-1. Study and control of these protein-protein interactions in vivo can be greatly promoted by introducing intracellular reagents that mimic EGFR functions. Here, we showed that a synthetic phosphopeptide encompassing the autophosphorylation site 1068 of EGFR formed a complex with endogenous Grb2 after this peptide was delivered into intact cells by a cell-permeable peptide import technique. Consequently, this intracellular peptide inhibited EGF-induced EGFR/Grb2 associations but not EGFR/Shc or Shc/Grb2 associations. Peptide-mediated disruption of the EGF/Grb2/Sos-1 cascade led to reduced Ras activation and mitogen-activated protein kinase activation. These results indicate that the binding of Grb2 to the phosphorylated Tyr-1068 of EGFR is crucial to the EGF-induced Ras/mitogen-activated protein kinase signaling pathway. The application of cell-permeable peptides to this study demonstrates a useful biochemical tool to probe and control various intracellular processes involved in signal transduction and gene transcription. |
| Databáze: | OpenAIRE |
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