Antiphosphotyrosine antibodies modulate insulin receptor kinase activity and insulin action.
Insulin elicits the autophosphorylation of the beta-subunit of its receptor on tyrosine residues: this effect appears to be the earliest post-binding event involved in insulin action. In the present study we have raised highly specific antibodies to phosphotyrosine residues, and we have taken advantage of these antibodies to further evaluate the role of the insulin receptor tyrosine kinase in the generation of insulin's biological responses. Using a cell-free phosphorylation assay, we show here that these antibodies increase the tyrosine kinase activity of the receptor, and its phosphorylation on tyrosine residues. In contrast, the antibodies do not interfere with dephosphorylation of the insulin receptor. Introduction of the same antibodies in living Fao hepatoma cells enhances the effect of insulin on both glucose transport and aminoacid uptake. As a whole our data indicate that the insulin receptor kinase is involved in the generation of an early (glucose transport) and late (aminoacid uptake) response to insulin. Further, conformational changes in phosphotyrosine containing domains of the insulin receptor appear to modulate insulin's biological effects. Finally, the injection of antibodies in intact cells provides us with a novel and promising tool to search for cellular substrates for the insulin receptor tyrosine kinase.
Ballotti
R
R
Institut National de la Santé et de la Recherche Médicale U 145, Faculté de Médecine, Nice, France.
Scimeca
J C
JC
Kowalski
A
A
Van Obberghen
E
E
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D000906:Antibodies; D000918:Antibody Specificity; D001692:Biological Transport; D002474:Cell-Free System; D005947:Glucose / Q000378:metabolism; D007328:Insulin / Q000378:metabolism*; D008114:Liver Neoplasms, Experimental; D010766:Phosphorylation; D019000:Phosphotyrosine; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D051381:Rats; D011972:Receptor, Insulin; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000276:immunology / Q000378:metabolism
D000596:Amino Acids; D000906:Antibodies; D007328:Insulin; D019000:Phosphotyrosine; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases; D011972:Receptor, Insulin; D005947:Glucose