cap alpha. -transforming growth factor secreted by untransformed bovine anterior pituitary cells in culture. II. Identification using a sequence-specific monoclonal antibody
Untransformed bovine anterior pituitary cells cultured in serum-free defined medium secrete an epidermal growth factor (EGF)-like peptide with an amino acid composition similar to rat or human ..cap alpha..-transforming growth factor (..cap alpha..TGF). To further characterize the bovine pituitary ..cap alpha..TGF, it was compared to a human ..cap alpha..TGF partially purified from the conditioned medium of a human melanoma cell line. An anti-..cap alpha..TGF monoclonal antibody, MF9, was produced from hybridomas derived from mice immunized with a 17-residue synthetic peptide corresponding to the carboxyl-terminal sequence of rat ..cap alpha..TGF. The hybridoma supernatants were initially screened for the ability to immunoprecipitate /sup 125/I-peptide and then tested for recognition of human ..cap alpha..TGF. Only 2 of 36 antipeptide antibodies recognized the native ..cap alpha..TGF. The binding of /sup 125/I-peptide to MF9 was displaced by human ..cap alpha..TGF but not by EGF. Bovine pituitary ..cap alpha..TGF also displaced the binding of /sup 125/I-peptide to MF9 in a similar manner to human ..cap alpha..TGF. Both iodinated human and bovine pituitary ..cap alpha..TGF were immunoprecipitated by MF9 whereas /sup 125/I-EGF was not. Tryptic digests of both /sup 125/I-..cap alpha..TGFs chromatographed to give a single, indistinguishable peak of iodinated material on a reverse-phase C/sub 18/ high performance liquid chromatography column when eluted with two different solvent systems, suggesting the generation of a single and identical tyrosine-containing tryptic peptide from both ..cap alpha..TGFs. The comparisons of the bovine pituitary and human melanoma ..cap alpha..TGF using a sequence-specific monoclonal antibody and peptide mapping suggest that these ..cap alpha..TGFs are related and that ..cap alpha..TGF production is not limited to transformed or fetal sources.
- Research Organization:
- Univ. of Toronto, Ontario
- OSTI ID:
- 6845152
- Journal Information:
- J. Biol. Chem.; (United States), Vol. 261:31
- Country of Publication:
- United States
- Language:
- English
Similar Records
Human transforming growth factor type. cap alpha. coding sequence is not a directed-acting oncogene when overexpressed in NIH 3T3 cells
Synthetic. cap alpha. subunit peptide 125-147 of human nicotinic acetylcholine receptor induces antibodies to native receptor
Related Subjects
59 BASIC BIOLOGICAL SCIENCES
PITUITARY GLAND
RADIORECEPTOR ASSAY
PROTEINS
AMINO ACID SEQUENCE
IMMUNOLOGY
ANIMAL CELLS
CATTLE
GROWTH
HYBRIDOMAS
IODINE 125
LIQUID COLUMN CHROMATOGRAPHY
MONOCLONAL ANTIBODIES
PRECIPITATION
TYROSINE
AMINO ACIDS
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BODY
CARBOXYLIC ACIDS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
ELECTRON CAPTURE RADIOISOTOPES
ENDOCRINE GLANDS
GLANDS
HYDROXY ACIDS
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
MAMMALS
MOLECULAR STRUCTURE
NUCLEI
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
RADIOISOTOPES
RUMINANTS
SEPARATION PROCESSES
TRACER TECHNIQUES
VERTEBRATES
550601* - Medicine- Unsealed Radionuclides in Diagnostics
550201 - Biochemistry- Tracer Techniques