Biosynthesis and secretion of follicle-stimulating hormone beta-subunit from ovine pituitary cultures: effect of 17 beta-estradiol treatment.
An assay was developed to detect tritium-labeled ovine FSH beta-subunit [( 3H]oFSH beta) secreted from primary ovine pituitary cultures. This procedure used affinity-enriched antibodies raised against reduced and carbamylmethylated oFSH beta (RCM-oFSH beta) in a two-cycle immunoextraction procedure. A discrete species with an apparent mol wt of 21,000 was detected in sodium dodecyl sulfate electrophoretic patterns of immunoextracts from culture medium. This species was identified as RCM-[3H]oFSH beta by its comigration with highly purified RCM-oFSH beta, its reduction in culture media after cultures were treated with 17 beta-estradiol, which normally decreases radioimmunoassayable oFSH; and its displacement from the extracting antibodies by excess unlabeled RCM-oFSH beta. The assay was used in a pulse-chase study to determine that [3H]oFSH beta is secreted within 1-2 h of its synthesis. Prior treatment of cultures with 17 beta-estradiol did not change this timing of secretion.
D000818:Animals; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D004958:Estradiol / Q000494:pharmacology*; D005640:Follicle Stimulating Hormone / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D037201:Follicle Stimulating Hormone, beta Subunit; D007158:Immunologic Techniques; D007460:Iodoacetamide; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D010902:Pituitary Gland / Q000187:drug effects / Q000378:metabolism*; D012756:Sheep; D014316:Tritium
D037201:Follicle Stimulating Hormone, beta Subunit; D010446:Peptide Fragments; D014316:Tritium; D004958:Estradiol; D005640:Follicle Stimulating Hormone; D007460:Iodoacetamide