Evidence for autocrine activation of a tyrosine kinase in a human gastric carcinoma cell line.
Phosphotyrosine (P-Tyr) antibodies have been used to identify the phosphorylated forms of growth factor receptors and oncogene-coded tyrosine kinases. Western blot analysis of a gastric carcinoma cell line with P-Tyr antibodies revealed a tyrosine-phosphorylated protein of Mr 145,000 (P145). In addition, in vitro phosphorylation with (gamma-32P)ATP or P-Tyr immunoprecipitates of the same cells resulted in labelling of this protein on tyrosine. P145 appears to be a transmembrane glycoprotein, with features suggestive of a growth factor receptor. However, the in vivo or in vitro addition of known growth factors did not affect P145 tyrosine phosphorylation. We now report that P145 is rapidly dephosphorylated in vivo when cells are exposed to low pH, a condition known to dissociate ligands from their receptors. The addition of serum-free medium, conditioned by the gastric carcinoma cells, fully restores the tyrosine phosphorylation lost with acid treatment. These data suggest that the activity responsible for P145 phosphorylation on tyrosine, whether intrinsic to P145 itself or due to an associated kinase, is stimulated by a factor secreted by the tumor cells themselves.
Giordano
S
S
Department of Biomedical Sciences and Oncology, University of Torino Medical School, Italy.
Di Renzo
M F
MF
Narsimhan
R P
RP
Tamagnone
L
L
Gerbaudo
E V
EV
Chiadó-Piat
L
L
Comoglio
P M
PM
D015153:Blotting, Western; D002460:Cell Line; D003470:Culture Media; D004789:Enzyme Activation; D006801:Humans; D009363:Neoplasm Proteins / Q000302:isolation & purification; D010750:Phosphoproteins / Q000302:isolation & purification; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D013274:Stomach Neoplasms / Q000201:enzymology*
D003470:Culture Media; D009363:Neoplasm Proteins; D010750:Phosphoproteins; D011505:Protein-Tyrosine Kinases