Cell-free translation of messenger RNA from bovine submaxillary glands and identification of the apomucin
This study was undertaken to identify and characterize the apoprotein of bovine submaxillary mucin (BSM). Purified preparations of BSM were deglycosylated by treatment with either anhydrous hydrogen fluoride or trifluoromethane-sulfonic acid. The amino acid compositions of the deglycosylated and native BSM were similar indicating that chemical deglycosylation did not cause significant degradation of the protein. Messenger RNA was isolated from bovine submaxillary glands by extraction of total RNA followed by chromatography on oligo(dT)-cellulose. The Poly A/sup +/ mRNA was translated in a rabbit reticulocyte cell-free translation system using either (/sup 35/S)methionine, (/sup 3/H)leucine or (/sup 3/H)threonine, as label, and the translation products analyzed by SDS-PAGE. The apoprotein of BSM was identified among the translation products by its immunoprecipitation with a specific polyclonal rabbit antiserum prepared against deglycosylated BSM. A product of about 65 kDA was precipitated with the antibody in the absence but not in the presence of deglycosylated BSM. Thus, it can be concluded that the primary translation product of the BSM gene is a 65 kDA protein. It is of interest that enzymatically deglycosylated ovine submaxillary mucin was found to have a molecular weight of 58 kDA.
- Research Organization:
- Penn State Univ., Hershey
- OSTI ID:
- 7108758
- Report Number(s):
- CONF-8606151-; TRN: 86-039030
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:6; Conference: 76. annual meeting of the Federation of American Society for Experimental Biology, Washington, DC, USA, 8 Jun 1986
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
MESSENGER-RNA
LABELLING
PROTEINS
CHEMICAL COMPOSITION
ANTIBODIES
CATTLE
CHROMATOGRAPHY
ELECTROPHORESIS
GLANDS
LEUCINE
METHIONINE
MOLECULAR WEIGHT
SULFUR 35
THREONINE
TRACER TECHNIQUES
TRITIUM COMPOUNDS
AMINO ACIDS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBOXYLIC ACIDS
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
DRUGS
EVEN-ODD NUCLEI
HYDROXY ACIDS
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
NUCLEI
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
RADIOISOTOPES
RNA
RUMINANTS
SEPARATION PROCESSES
SULFUR ISOTOPES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques