Sequential processing reactions in the formation of hormone amides.
The substrate specificity of an enzyme with amidating activity, present in porcine pituitary, was investigated by examining its ability to convert the synthetic peptides D-Tyr-Val-Gly and D-Tyr-Val-Gly-Lys-Arg to the dipeptide amide D-Tyr-Val-CONH2. The purified enzyme catalysed the amidation reaction with the tripeptide but did not accept the pentapeptide as a substrate. With the mixture of enzymes present in a membrane fraction from porcine pituitary or the enzymes in a secretory granule fraction, both the tripeptide and pentapeptide substrates gave rise to D-Tyr-Val amide; the formation of dipeptide amide from the pentapeptide, however, involved a latency period after which amidation occurred at a similar rate with the two substrates. Evidence was obtained that arginine and lysine were released from the C terminus of the pentapeptide before amidation took place since the rate of formation of dipeptide amide was reduced at pH values that were compatible with amidation but unfavourable to the action of carboxypeptidase H. In addition formation of the dipeptide amide from the pentapeptide was blocked by guanidinoethylmercaptosuccinic acid and glycylarginine, which are inhibitors of carboxypeptidase enzymes. The experiments demonstrate that removal of basic residues from the C terminus of a peptide and amidation at C-terminal glycine are reactions that take place consecutively. These prohormone-processing reactions, which are intrinsic to the formation of hormone amides, did not synergise.
D000581:Amidohydrolases / Q000378:metabolism*; D000818:Animals; D003594:Cytoplasmic Granules / Q000201:enzymology; D006728:Hormones / Q000378:metabolism*; D009842:Oligopeptides / Q000378:metabolism; D010902:Pituitary Gland / Q000201:enzymology*; D011498:Protein Precursors / Q000378:metabolism; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives; D013379:Substrate Specificity; D013552:Swine; D013973:Thyrotropin-Releasing Hormone / Q000378:metabolism
D006728:Hormones; D009842:Oligopeptides; D011498:Protein Precursors; D013973:Thyrotropin-Releasing Hormone; C046418:pro-thyrotropin releasing hormone; D000581:Amidohydrolases; D011761:Pyrrolidonecarboxylic Acid