Characterization of proopiocortin converting activity in rat anterior pituitary secretory granules.
Lysates from purified secretory granules of rat anterior pituitary glands were incubated with [3H]phenylalanine or [3H]arginine-labeled toad proopiocortin. The processed products formed were identified by immunoprecipitation with ACTH and beta-endorphin antisera, and by comigration with known markers on acid-urea polyacrylamide gels. Proopiocortin was cleaved by the secretory granule lysate primarily to 21,000 mol wt ACTH, 13,000 mol wt ACTH, 16,000 mol wt NH2-terminal glycopeptide, beta-lipotropin, a beta-endorphin-like peptide, and beta-endorphin. Characterization of the anterior pituitary proopicortin-converting activity shows that it: (1) cleaves specifically at the peptide bond on the carboxy side of the lysine-arginine residues of proopiocortin, (2) has a pH optimum in the acidic range, (3) is present in membrane and soluble fractions of the granule lysate, and (4) is inhibited by leupeptin, pepstatin A, and 2,2' dithiodipyridine, but not by p-chloromercuribenzoate, diisopropyl fluorophosphate, N alpha-p-tosyl-L-lysine chloromethyl ketone hydrochloride, chloroquine, L-1-tosylamide-2-phenylethyl-chloromethyl ketone, or EDTA.
D000324:Adrenocorticotropic Hormone / Q000032:analysis; D000818:Animals; D043424:Carboxypeptidase B; D002268:Carboxypeptidases; D003594:Cytoplasmic Granules / Q000201:enzymology*; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D005260:Female; D005966:Glucuronidase / Q000032:analysis; D007425:Intracellular Membranes / Q000201:enzymology; D007531:Isoflurophate / Q000494:pharmacology; D007700:Kinetics; D008995:Monoamine Oxidase / Q000032:analysis; D010903:Pituitary Gland, Anterior / Q000201:enzymology*; D043484:Proprotein Convertases; D051381:Rats; D011919:Rats, Inbred Strains; D014981:Xenopus
D007531:Isoflurophate; D000324:Adrenocorticotropic Hormone; D008995:Monoamine Oxidase; D005966:Glucuronidase; D002268:Carboxypeptidases; D010450:Endopeptidases; D043424:Carboxypeptidase B; D043484:Proprotein Convertases