The extracellular domain of p185/neu is released from the surface of human breast carcinoma cells, SK-BR-3.
The human breast carcinoma cell line SK-BR-3, expresses the neu oncogene product, p185, which is a receptor tyrosine kinase. Using a double monoclonal antibody capture enzyme-linked immunosorbent assay for p185, activity was detected in conditioned media from cultures of SK-BR-3 cells. Two monoclonal antibodies specific for the extracellular domain of p185/neu immunoprecipitated a protein with a molecular mass of approximately 105 kDa. p105 was further shown to compete with p185 for binding to monoclonal antibodies and pulse-chase experiments indicate that it was generated by post-translational processing. Peptide maps showed that p105 and p185 are related polypeptides. Since p105 is close to the predicted size for the extracellular domain of p185/neu, we propose that SK-BR-3 cells specifically process and release this portion of the receptor into the medium. The release of the extracellular domain may have implications in oncogenesis and its detection could prove useful as a cancer diagnostic.
Zabrecky
J R
JR
Applied bioTechnology, Inc., Cambridge, Massachusetts 02142.
Lam
T
T
McKenzie
S J
SJ
Carney
W
W
D000230:Adenocarcinoma / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000276:immunology; D001943:Breast Neoplasms / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D066298:In Vitro Techniques; D008970:Molecular Weight; D010446:Peptide Fragments / Q000378:metabolism; D010449:Peptide Mapping; D011233:Precipitin Tests; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D018719:Erb-b2 Receptor Tyrosine Kinases; D012995:Solubility; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D011518:Proto-Oncogene Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases