The presence of c-erbB-2 gene product-related protein in culture medium conditioned by breast cancer cell line SK-BR-3
Abstract
The Mr 185,000 glycoprotein encoded by human c-erbB-2/neu/HER2 gene, termed c-erbB-2 gene product, shows a close structural similarity with epidermal growth factor receptor and is now regarded to be a growth factor receptor for an as yet unidentified ligand. Abundant c-erbB-2 mRNA was demonstrated by Northern blot studies in the human breast cancer cell line SK-BR-3. Cellular radiolabeling experiments followed by immunoprecipitation with three different anti-c-erbB-2 gene product antibodies, recognizing extracellular domain, kinase domain, and carboxyl-terminal portion, respectively, demonstrated the production of a large amount of c-erbB-2 gene product which had the capacity to be phosphorylated. Immunization of mice with concentrated culture medium conditioned by SK-BR-3 cells always generated antibodies against c-erbB-2 gene product, demonstrating that this culture medium contained substance(s) immunologically indistinguishable from c-erbB-2 gene product. This observation was supported by the successful development of a monoclonal antibody against c-erbB-2 gene product, GFD-OA-p185-1, by immunizing mice with this culture medium. The biochemical nature of the substance(s) present in the culture medium was further characterized. When the culture medium conditioned by (35S)cysteine-labeled SK-BR-3 cells was immunoprecipitated by three different anti-c-erbB-2 gene product antibodies, only the antibody recognizing extracellular domain precipitated the (35S)-labeled protein with a molecular weight of 110,000, namelymore »
- Authors:
-
- National Cancer Center Research Institute, Tokyo (Japan)
- Publication Date:
- OSTI Identifier:
- 5857656
- Resource Type:
- Journal Article
- Journal Name:
- Cell Growth and Differentation; (USA)
- Additional Journal Information:
- Journal Volume: 1:12; Journal ID: ISSN 1044-9523
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; GLYCOPROTEINS; GENES; GROWTH FACTORS; RECEPTORS; CULTURE MEDIA; CYSTEINE; IMMUNOASSAY; MAMMARY GLANDS; MAN; MICE; MOLECULAR STRUCTURE; MOLECULAR WEIGHT; MONOCLONAL ANTIBODIES; NEOPLASMS; ONCOGENES; PHOSPHOPROTEINS; PHOSPHORYLATION; SULFUR 35; TRACER TECHNIQUES; TUMOR CELLS; AMINO ACIDS; ANIMAL CELLS; ANIMALS; ANTIBODIES; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; BIOASSAY; BODY; CARBOXYLIC ACIDS; CHEMICAL REACTIONS; DAYS LIVING RADIOISOTOPES; DISEASES; EVEN-ODD NUCLEI; GLANDS; ISOTOPE APPLICATIONS; ISOTOPES; LIGHT NUCLEI; MAMMALS; MEMBRANE PROTEINS; MITOGENS; NUCLEI; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC SULFUR COMPOUNDS; ORGANS; PRIMATES; PROTEINS; RADIOISOTOPES; RODENTS; SULFUR ISOTOPES; THIOLS; VERTEBRATES; 550201* - Biochemistry- Tracer Techniques; 550901 - Pathology- Tracer Techniques
Citation Formats
Alper, O, Yamaguchi, K, Hitomi, J, Honda, S, Matsushima, T, and Abe, K. The presence of c-erbB-2 gene product-related protein in culture medium conditioned by breast cancer cell line SK-BR-3. United States: N. p., 1990.
Web.
Alper, O, Yamaguchi, K, Hitomi, J, Honda, S, Matsushima, T, & Abe, K. The presence of c-erbB-2 gene product-related protein in culture medium conditioned by breast cancer cell line SK-BR-3. United States.
Alper, O, Yamaguchi, K, Hitomi, J, Honda, S, Matsushima, T, and Abe, K. 1990.
"The presence of c-erbB-2 gene product-related protein in culture medium conditioned by breast cancer cell line SK-BR-3". United States.
@article{osti_5857656,
title = {The presence of c-erbB-2 gene product-related protein in culture medium conditioned by breast cancer cell line SK-BR-3},
author = {Alper, O and Yamaguchi, K and Hitomi, J and Honda, S and Matsushima, T and Abe, K},
abstractNote = {The Mr 185,000 glycoprotein encoded by human c-erbB-2/neu/HER2 gene, termed c-erbB-2 gene product, shows a close structural similarity with epidermal growth factor receptor and is now regarded to be a growth factor receptor for an as yet unidentified ligand. Abundant c-erbB-2 mRNA was demonstrated by Northern blot studies in the human breast cancer cell line SK-BR-3. Cellular radiolabeling experiments followed by immunoprecipitation with three different anti-c-erbB-2 gene product antibodies, recognizing extracellular domain, kinase domain, and carboxyl-terminal portion, respectively, demonstrated the production of a large amount of c-erbB-2 gene product which had the capacity to be phosphorylated. Immunization of mice with concentrated culture medium conditioned by SK-BR-3 cells always generated antibodies against c-erbB-2 gene product, demonstrating that this culture medium contained substance(s) immunologically indistinguishable from c-erbB-2 gene product. This observation was supported by the successful development of a monoclonal antibody against c-erbB-2 gene product, GFD-OA-p185-1, by immunizing mice with this culture medium. The biochemical nature of the substance(s) present in the culture medium was further characterized. When the culture medium conditioned by (35S)cysteine-labeled SK-BR-3 cells was immunoprecipitated by three different anti-c-erbB-2 gene product antibodies, only the antibody recognizing extracellular domain precipitated the (35S)-labeled protein with a molecular weight of 110,000, namely p110. The newly developed monoclonal antibody also immunoprecipitated this protein.},
doi = {},
url = {https://www.osti.gov/biblio/5857656},
journal = {Cell Growth and Differentation; (USA)},
issn = {1044-9523},
number = ,
volume = 1:12,
place = {United States},
year = {Sat Dec 01 00:00:00 EST 1990},
month = {Sat Dec 01 00:00:00 EST 1990}
}