Synthetic phosphopeptide immunogens yield activation-specific antibodies to the c-erbB-2 receptor.
We inoculated rabbits with synthetic phosphopeptides, duplicating a major autophosphorylation site of the c-erbB-2 protooncogene product. The rabbits produced antisera that, after reverse immunoaffinity purification, selectively recognize the erbB-2 protein in its enzymatically active configuration. These anti-phosphopeptide antisera identify a subset of erbB-2-positive human cell lines wherein the protein is constitutively active as a tyrosine kinase. Synthetic phosphopeptides incorporating informative protein phosphorylation sites may prove useful for generating antibodies that indicate the activation state of additional tyrosine kinases and perhaps other proteins phosphorylated on serine and threonine residues.
Division of Cell and Molecular Biology, Dana-Farber Cancer Institute, Boston, MA.
R J
RJ
Epstein
B J
BJ
Druker
T M
TM
Roberts
C D
CD
Stiles
D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification; D000936:Antigen-Antibody Complex; D002460:Cell Line; D005260:Female; D006801:Humans; D051379:Mice; D010748:Phosphopeptides / Q000138:chemical synthesis / Q000276:immunology*; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011519:Proto-Oncogenes; D051381:Rats; D018719:Erb-b2 Receptor Tyrosine Kinases; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000936:Antigen-Antibody Complex; D010748:Phosphopeptides; D011505:Protein-Tyrosine Kinases; D011518:Proto-Oncogene Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D013762:Tetrahydrofolate Dehydrogenase