cap alpha. -transforming growth factor secreted by untransformed bovine anterior pituitary cells in culture. II. Identification using a sequence-specific monoclonal antibody

Journal Article · Wed Nov 05 04:00:00 EST 1986 · J. Biol. Chem.; (United States)

OSTI ID:6845152

Kobrin, M S; Samsoondar, J; Kudlow, J E

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), Journal Name: J. Biol. Chem.; (United States) Vol. 261:31; ISSN JBCHA
• Country of Publication: United States
• Language: English

Related Subjects

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59 BASIC BIOLOGICAL SCIENCES
62 RADIOLOGY AND NUCLEAR MEDICINE
AMINO ACID SEQUENCE
AMINO ACIDS
ANIMAL CELLS
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BODY
CARBOXYLIC ACIDS
CATTLE
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
ELECTRON CAPTURE RADIOISOTOPES
ENDOCRINE GLANDS
GLANDS
GROWTH
HYBRIDOMAS
HYDROXY ACIDS
IMMUNOLOGY
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LIQUID COLUMN CHROMATOGRAPHY
MAMMALS
MOLECULAR STRUCTURE
MONOCLONAL ANTIBODIES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PITUITARY GLAND
PRECIPITATION
PROTEINS
RADIOISOTOPES
RADIORECEPTOR ASSAY
RUMINANTS
SEPARATION PROCESSES
TRACER TECHNIQUES
TYROSINE
VERTEBRATES