Site-specific association of c-Src with epidermal growth factor receptor in A431 cells.
We have examined the interaction between c-Src and epidermal growth factor (EGF) receptor in A431 cells. c-Src was found exclusively in the Triton X-100-solubilized particulate fraction and activated up to 3-fold within 1 min after EGF treatment of the cells. Association between c-Src and EGF receptor was detected by immunoprecipitation of c-Src followed by immunoblotting with anti-EGF receptor antibody. The c-Src-EGF receptor complex was found in both EGF-treated and untreated cells, but an augmented complex formation was observed in EGF-treated cells. We have isolated the complex by DEAE-cellulose column chromatography and found that a site-specific anti-c-Src antibody, which was raised against a synthetic peptide corresponding to residues 413 to 431 of human c-Src, did not recognize the c-Src protein in the complex, while other c-Src-specific antibodies tested did. Incubation of the complex with this synthetic peptide resulted in a partial dissociation of the complex. These results suggest that the specific region of c-Src is involved in the association with EGF receptor.
Sato
K
K
Laboratory of Molecular Biology, Kobe University, Japan.
Sato
A
A
Aoto
M
M
Fukami
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001665:Binding Sites; D002294:Carcinoma, Squamous Cell; D002460:Cell Line; D002848:Chromatography, DEAE-Cellulose; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004815:Epidermal Growth Factor / Q000494:pharmacology; D066246:ErbB Receptors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D008969:Molecular Sequence Data; D017830:Octoxynol; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000906:Antibodies; D010455:Peptides; D004815:Epidermal Growth Factor; D017830:Octoxynol; D066246:ErbB Receptors; D016392:Proto-Oncogene Proteins pp60(c-src)