title: Amidation of joining peptide, a major pro-ACTH/endorphin-derived product peptide.
abstract: Based on sequence data, rat and mouse pro-adrenocorticotropin (ACTH)/endorphin could give rise to joining peptide, a short acidic peptide that could terminate with a glutamic acid alpha-amide. Rat and mouse pituitary cells were found to cleave the pro-ACTH/endorphin precursor at an -Arg-Arg- site to produce primarily joining peptide-sized material. The amounts of joining peptide were approximately equimolar to the other major pro-ACTH/endorphin-derived products. Using antisera specific for the COOH-terminal modifications of joining peptide and three analytical approaches which separate amidated from glycine-extended forms of joining peptide, it was found that most of the joining peptide in murine anterior and intermediate pituitary was amidated. Identification of the amidated and glycine-extended forms of joining peptide was confirmed by amino acid analysis of the purified molecules. When anterior pituitary corticotrope tumor cells were grown in culture medium lacking ascorbate, there was no detectable ascorbate in the cells; nevertheless, a significant fraction of the joining peptide produced was alpha-amidated, indicating that production of alpha-amidated product was not totally dependent on ascorbate. The amidation state of the joining peptide produced by mouse corticotrope tumor cells was responsive to added ascorbate. Cells grown in medium containing ascorbic acid at the levels found in plasma concentrated the ascorbate to the levels normally found in pituitary tissue, and nearly all of the joining peptide produced was alpha-amidated. The amidation state of secreted joining peptide mirrored the amidation state of the joining peptide in the cells.
mesh_terms: D000577:Amides; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D002460:Cell Line; D002850:Chromatography, Gel; D008297:Male; D051379:Mice; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis* / Q000302:isolation & purification; D010902:Pituitary Gland / Q000032:analysis*; D010903:Pituitary Gland, Anterior / Q000032:analysis; D010911:Pituitary Neoplasms / Q000032:analysis*; D011333:Pro-Opiomelanocortin / Q000032:analysis* / Q000302:isolation & purification; D011499:Protein Processing, Post-Translational; D051381:Rats
chemical_list: D000577:Amides; D000596:Amino Acids; D010446:Peptide Fragments; C049531:pro-opiomelanocortin joining peptide(77-97); D011333:Pro-Opiomelanocortin; D001205:Ascorbic Acid