Document Detail

Purification and characterization of the neu/erb B2 ligand-growth factor from bovine kidney.
MedLine Citation:
PMID:  1350785     Owner:  NLM     Status:  MEDLINE    
A neu/erb B2 ligand growth factor (NEL-GF) was purified to homogeneity from bovine kidney by a procedure involving ammonium sulfate fractionation (35-70% saturation) followed by sequential column chromatography on DEAE-cellulose (DE52), Sulfadex (sulfated Sephadex G-50), heparin-Sepharose 4B, and Superdex 75 (fast protein liquid chromatography). NEL-GF was found to be a 25-kDa polypeptide according to the analysis by gel filtration on Superdex 75 and 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. NEL-GF stimulated the tyrosine-specific autophosphorylation of the neu/erb B2 gene product purified by immunoabsorbent and tyrosine-specific phosphorylation of the neu/erb B2 gene product in intact dihydrofolate reductase (DHFR/G-8 cells (NIH 3T3 cells transfected with rat c-neu). NEL-GF also down-regulated the cell surface neu/erb B2 gene product in DHFR/G-8 cells. NEL-GF was mitogenic toward NIH 3T3 cells, DHFR/G-8 cells, A431 cells (human epidermoid carcinoma cells), and SK-BR-3 cells (human breast carcinoma cells) but inactive toward bovine aorta endothelial cells. NEL-GF was sensitive to 0.1% trifluoroacetic acid but resistant to 5% beta-mercaptoethanol and appeared to be distinct from a neu protein-specific activating factor (Davis, J. G., Hamuro, J., Shim, C. Y., Samanta, A., Greene, M. I., and Dobashi, K. (1991) Biochem. Biophys. Res. Commun. 179, 1536-1542) and a 30-kDa glycoprotein which competed with a monoclonal antibody for binding to the neu/erb B2 gene product (Lupu, R., Colomer, R., Zugmaier, G., Sarup, J., Shepard, M., Slamon, D., and Lippman, M. E. (1990) Science 249, 1552-1555).
S S Huang; J S Huang
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  267     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1992 Jun 
Date Detail:
Created Date:  1992-07-06     Completed Date:  1992-07-06     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  11508-12     Citation Subset:  IM    
Department of Biochemistry and Molecular Biology, St. Louis University School of Medicine, Missouri 63104.
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MeSH Terms
3T3 Cells
Cells, Cultured
Chromatography, Gel
Electrophoresis, Polyacrylamide Gel
Endothelium, Vascular / cytology,  metabolism
Kidney / metabolism*
Protein-Tyrosine Kinases / metabolism*
Proto-Oncogene Proteins / isolation & purification*,  metabolism
Receptor, erbB-2
Tetrahydrofolate Dehydrogenase / metabolism
Tumor Cells, Cultured
Grant Support
Reg. No./Substance:
0/Proto-Oncogene Proteins; EC Dehydrogenase; EC Kinases; EC, erbB-2

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