Disulfide-linked and noncovalent dimers of p185HER-2 in human breast carcinoma cells.
Enhanced levels of disulfide-linked dimers of the neu oncogene product have been suggested to be associated with the transformed state [Weiner DB, Liu J, Cohen JA, Williams WV, Greene MI: Nature 338:230-231, (1989)]. We, therefore, investigated the properties of the dimeric forms of p185HER-2/neu from the human breast carcinoma cell line, SK-BR-3. We found disulfide-linked dimers as well as noncovalently associated dimers that were detected by cross-linking with bis(sulfosuccinimidyl) suberate (BS3). However, the disulfide-linked dimers did not exist in intact cells, since they were eliminated when the cells were lysed in the presence of the alkylating agent, sodium iodoacetate. Moreover, the disulfide-linked dimeric molecules were not the activated form of p185HER-2 since they incorporated about the same level of phosphate in an in vitro kinase reaction as the monomeric molecules. In contrast, the noncovalent dimers appeared to be present on the surface of intact cells and were phosphorylated at levels at least tenfold higher than monomers in an in vitro kinase reaction.
Lin
Y J
YJ
Department of Biochemistry and Molecular Biology, Oregon Health Sciences University, Portland 97201.
Christianson
T A
TA
Clinton
G M
GM
D000477:Alkylating Agents / Q000494:pharmacology; D000906:Antibodies / Q000276:immunology; D001943:Breast Neoplasms; D004220:Disulfides / Q000737:chemistry*; D006801:Humans; D007461:Iodoacetates / Q000494:pharmacology; D019807:Iodoacetic Acid; D009856:Oncogene Proteins, Viral / Q000737:chemistry* / Q000378:metabolism; D010766:Phosphorylation; D019000:Phosphotyrosine; D011233:Precipitin Tests; D018719:Erb-b2 Receptor Tyrosine Kinases; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000031:analogs & derivatives / Q000276:immunology
D000477:Alkylating Agents; D000906:Antibodies; D004220:Disulfides; D007461:Iodoacetates; D019807:Iodoacetic Acid; D009856:Oncogene Proteins, Viral; D019000:Phosphotyrosine; D018719:Erb-b2 Receptor Tyrosine Kinases; D014443:Tyrosine