Numerous proteins phosphorylated on tyrosine and enhanced tyrosine kinase activities in vanadate-treated NIH 3T3 fibroblasts.
A monoclonal antibody that can immunoprecipitate proteins containing phosphotyrosine has been isolated and characterized. To identify proteins that can act as substrates for tyrosine kinases in intact cells, extracts of phosphate-labeled NIH cells that had been treated with the phosphotyrosyl phosphatase inhibitor, vanadate, were precipitated with the antibody, and the immunoprecipitates were analyzed by two-dimensional gel electrophoresis. Numerous proteins were specifically precipitated from vanadate-treated NIH 3T3 cells by the antibody. The high level of phosphotyrosine detected in vanadate-treated cells is presumably primarily due to phosphatase inhibition, but approx. 2-fold increased tyrosine kinase activities were also detected in extracts of the cells after treatment with vanadate. The enhanced tyrosine kinase activity may contribute to the generation of the transformed phenotype seen in response to treatment with vanadate.
Klarlund
J K
JK
Fibiger Institute, Danish Cancer Society, Copenhagen, Denmark.
Latini
S
S
Forchhammer
J
J
D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D005347:Fibroblasts / Q000201:enzymology / Q000378:metabolism*; D006241:Haptens / Q000276:immunology; D006433:Hemocyanins / Q000276:immunology; D051379:Mice; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism*; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000276:immunology / Q000378:metabolism*; D014638:Vanadates
D000911:Antibodies, Monoclonal; D006241:Haptens; D010750:Phosphoproteins; D014638:Vanadates; D014443:Tyrosine; D006433:Hemocyanins; D011505:Protein-Tyrosine Kinases