Receptor-induced phosphorylation by mammary-derived growth factor 1 in mammary epithelial cell lines.
Previous work has shown that a mammary-derived growth factor (MDGF1), a human milk-derived, acidic, 62-kDa, N-glycosylated growth factor binds to cell surface receptors and stimulates proliferation of mammary epithelial cells. An 18-amino acid N-terminal partial sequence of the factor did not show any homology to other known growth factors or proteins. Using polyclonal antiserum raised against the synthetic peptide, we demonstrated that conditioned medium prepared from human breast cancer cell lines contains the factor. The antibody could adsorb the biological activity of the factor present in the conditioned medium. Earlier experiments on receptor cross-linking indicated that the receptor was approximately 120-140 kDa. Since tyrosine phosphorylation plays a crucial role in cell proliferation and cell transformation, experiments were conducted to find out whether MDGF1 induces the appearance of phosphotyrosine in MDGF1-receptor-positive MDA-MB 468, MCF-7, and 184A1N4 cell lines compared to receptor-negative lines. Western blot analysis using monoclonal antiphosphotyrosine indicated that MDGF1 induces phosphotyrosine in a 180-185-kDa protein in MDGF1 receptor-positive cell lines. Phosphorylation was not blocked and phosphorylated proteins were not immunoprecipitated by an antibody directed against the binding site of the EGF receptor. Cell membrane fractionation demonstrated that phosphorylation induced by MDGF1 was membrane-associated. The nature of this 180-185-kDa protein and its possible relationship to the MDGF1 receptor are under investigation.
Bano
M
M
Lombardi Cancer Research Center, Georgetown University, Washington, D.C. 20007.
Worland
P
P
Kidwell
W R
WR
Lippman
M E
ME
Dickson
R B
RB
D015153:Blotting, Western; D001940:Breast / Q000166:cytology / Q000378:metabolism*; D001943:Breast Neoplasms / Q000378:metabolism*; D002460:Cell Line; D003432:Cross-Linking Reagents; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism; D006133:Growth Substances / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D009363:Neoplasm Proteins / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine; D011956:Receptors, Cell Surface / Q000378:metabolism*; D017978:Receptors, Growth Factor; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D003432:Cross-Linking Reagents; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; C062654:MDGF1 protein, human; D009363:Neoplasm Proteins; D011956:Receptors, Cell Surface; D017978:Receptors, Growth Factor; C045223:mammary-derived growth factor 1; D019000:Phosphotyrosine; D014443:Tyrosine; D066246:ErbB Receptors