Monoclonal Antibodies to Individual Tyrosine-Phosphorylated Protein Substrates of Oncogene-Encoded Tyrosine Kinases

• Published in: Proceedings of the National Academy of Sciences - PNAS 1990-05, Vol.87 (9), p.3328-3332
• Main Authors: Kanner, Steven B., Reynolds, Albert B., Vines, Richard R., Parsons, J. Thomas
• Format: Article
• Language: English
• Subjects: 550201 - Biochemistry- Tracer Techniques; AMINO ACIDS; Analytical, structural and metabolic biochemistry; ANIMAL CELLS; ANIMALS; ANTIBODIES; Antibodies, Monoclonal; Antigen-Antibody Complex; Antigens; BASIC BIOLOGICAL SCIENCES; BETA DECAY RADIOISOTOPES; BIOASSAY; Biological and medical sciences; BIRDS; CARBOXYLIC ACIDS; Cell Line; Cell lines; Cell Transformation, Neoplastic; CELL TRANSFORMATIONS; Cells, Cultured; CHEMICAL REACTIONS; Chick Embryo; CHICKENS; DAYS LIVING RADIOISOTOPES; ELECTRON CAPTURE RADIOISOTOPES; EMBRYONIC CELLS; ENZYME IMMUNOASSAY; Enzyme-Linked Immunosorbent Assay; ENZYMES; Enzymes and enzyme inhibitors; FOWL; Fundamental and applied biological sciences. Psychology; GENES; Humans; HYDROXY ACIDS; IMMUNOASSAY; Immunoblotting; Immunoglobulin G; Immunoglobulin M; Immunoprecipitation; INTERMEDIATE MASS NUCLEI; INTERNAL CONVERSION RADIOISOTOPES; IODINE 125; IODINE ISOTOPES; ISOTOPES; MICROORGANISMS; MONOCLONAL ANTIBODIES; NUCLEI; ODD-EVEN NUCLEI; Oncogene Protein pp60(v-src) - genetics; Oncogene Protein pp60(v-src) - metabolism; ONCOGENES; ONCOGENIC TRANSFORMATIONS; ORGANIC ACIDS; ORGANIC COMPOUNDS; PARASITES; Phosphoproteins; Phosphoproteins - analysis; PHOSPHORUS-GROUP TRANSFERASES; PHOSPHORYLATION; PHOSPHOTRANSFERASES; Protein-Tyrosine Kinases - genetics; Protein-Tyrosine Kinases - metabolism; Proteins; RADIOASSAY; RADIOISOTOPES; Spleen cells; SUBSTRATES; TRANSFERASES; TYROSINE; VERTEBRATES; VIRUSES
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.87.9.3328

Cellular transformation by oncogenic retro-viruses encoding protein tyrosine kinases coincides with the tyrosine-specific phosphorylation of multiple protein substrates. Previous studies have shown that tyrosine phosphorylation of a protein of 120 kDa, p120, correlated with src transformation in chicken embryo fibroblasts. Additionally, we previously identified two phosphotyrosine-containing cellular proteins, p130 and p110, that formed stable complexes with activated variants of pp60src, the src-encoded tyrosine kinase. To study transformation-relevant tyrosine kinase substrates, we have generated monoclonal antibodies to individual tyrosine phosphoproteins, including p130, p120, p110, and five additional phosphoproteins (p210, p125, p118, p85, and p185/p64). These antibodies detected several of the same tyrosine phosphoproteins in chicken embryo fibroblasts transformed by avian retroviruses Y73 and CT10, encoding the yes and crk oncogenes, respectively. Protein substrates in mouse, rat, hamster, and human cells overexpressing activated variants of chicken pp60srcwere also detected by several of the monoclonal antibodies.