Abstract
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.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Antibodies / immunology*
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Antibodies / isolation & purification
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Antigen-Antibody Complex
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Cell Line
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Female
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Humans
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Mice
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Phosphopeptides / chemical synthesis
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Phosphopeptides / immunology*
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Protein-Tyrosine Kinases / genetics
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Protein-Tyrosine Kinases / immunology*
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Protein-Tyrosine Kinases / metabolism
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Proto-Oncogene Proteins / genetics
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Proto-Oncogene Proteins / immunology*
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Proto-Oncogene Proteins / metabolism
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Proto-Oncogenes*
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Rats
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Receptor, ErbB-2
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Tetrahydrofolate Dehydrogenase / genetics
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Tetrahydrofolate Dehydrogenase / metabolism
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Transfection
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Tumor Cells, Cultured
Substances
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Antibodies
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Antigen-Antibody Complex
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Phosphopeptides
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Proto-Oncogene Proteins
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Tetrahydrofolate Dehydrogenase
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Protein-Tyrosine Kinases
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Receptor, ErbB-2