Processing, turnover and release of corticotropins, endorphins and melanotropin in the toad pituitary intermediate lobe.
The significance of glycosylation of the ACTH/alpha-MSH-endorphin precursor in the biosynthesis, processing and secretion of its peptide products was examined in the toad neurointermediate (intermediate - posterior) lobe, with the aid of a specific inhibitor of glycosylation, tunicamycin. Tunicamycin did not affect the synthesis of the precursor but prevented its glycosylation. In the presence of tunicamycin the precursor underwent rapid intracellular degradation. Precursor molecules that escaped complete degradation were processed to an ACTH molecule with approximately 19 000 molecular weight and to other atypical peptides, which were released. In vitro studies showed that trypsinization of the non-glycosylated precursor resulted in its random proteolysis while large forms of ACTH were cleaved from the glycosylated precursor. The results indicate that glycosylation of the ACTH/alpha-MSH-endorphin precursor may confer specific conformational properties upon the molecule, thus regulating its limited proteolysis. Turnover and release studies revealed two different pools of ACTH, beta-LPH and alpha-MSH-related peptides in the toad intermediate lobe. One pool contained ACTH, beta-LPH, alpha-MSH and beta-endorphin, which were rapidly synthesized and released, or degraded within 6 h of synthesis if their release was inhibited. The other pool was stored and was stable for at least 10 h, if prevented from being released. Peptides in this stored pool primarily included ACTH, alpha-MSH and beta-LPH; beta-endorphin was a minor component of this pool. The release from both pools of peptides was inhibited by dopamine, while the stored pool was selectively inhibited from release by L-isoprenaline (L-isoproterenol).
D000324:Adrenocorticotropic Hormone / Q000378:metabolism*; D000818:Animals; D004723:Endorphins / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism; D009074:Melanocyte-Stimulating Hormones / Q000378:metabolism*; D010902:Pituitary Gland / Q000187:drug effects / Q000378:metabolism*; D010908:Pituitary Hormones, Anterior / Q000378:metabolism*; D011333:Pro-Opiomelanocortin; D011498:Protein Precursors / Q000302:isolation & purification / Q000378:metabolism*; D014415:Tunicamycin / Q000494:pharmacology; D014981:Xenopus; D008083:beta-Lipotropin / Q000378:metabolism
D004723:Endorphins; D006023:Glycoproteins; D010908:Pituitary Hormones, Anterior; D011498:Protein Precursors; D014415:Tunicamycin; D011333:Pro-Opiomelanocortin; D000324:Adrenocorticotropic Hormone; D009074:Melanocyte-Stimulating Hormones; D008083:beta-Lipotropin