title: Evidence of glucagon biosynthesis involving protein intermediates in rat salivary glands.
abstract: In an attempt to determine the ability of rat submaxillary glands to synthesise glucagon via protein intermediates, isolated cells from these glands were incubated in vitro with 3H-L-tryptophan and the acid-ethanol extracts of the cells were purified on Bio-Gel P-30 columns. Aliquots of the eluates were incubated with a C-terminal glucagon antiserum (30K) and the radioactivity bound to the glucagon antibody appeared to be distributed among proteins of Molecular weight greater than 40.14 and 3.5 Kdaltons. A similar elution pattern was obtained in the presence of urea (7 mol/l) and guanidine hydrochloride (6 mol/l). To determine the molecular weight of the immunoreactive material eluting before the 3.5 Kdalton polypeptide, aliquots of the cell extracts were immunoprecipitated and analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Polypeptides of 125.8, 63.1, 42.6 and 14.4 Kdaltons were obtained. These polypeptides incorporate more radioactive tryptophan with increase in the time of incubation. Pulse-chase experiments with unlabelled tryptophan, cycloheximide-treatment of isolated cells and limited tryptic digestion of the larger glucagon immunoreactive component, transform it into a 3.5 Kdalton polypeptide with immunological characteristics indistinguishable from pancreatic glucagon. These results suggest that the larger molecule contains glucagon and thus may serve as a precursor or an intermediate of extrapancreatic glucagon biosynthesis.
mesh_terms: D000818:Animals; D003513:Cycloheximide / Q000494:pharmacology; D005260:Female; D005934:Glucagon / Q000096:biosynthesis*; D066298:In Vitro Techniques; D007700:Kinetics; D010452:Peptide Biosynthesis; D010446:Peptide Fragments / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D013363:Submandibular Gland / Q000187:drug effects / Q000378:metabolism*; D014316:Tritium
chemical_list: D010446:Peptide Fragments; D014316:Tritium; D005934:Glucagon; D003513:Cycloheximide