title: Peptidyl-glycine alpha-amidating monooxygenase is present in islet secretory granules of the anglerfish, Lophius americanus.
abstract: Anglerfish islet secretory granules have been examined for the presence of an enzyme which could perform C-terminal amidation of glucagon-like peptide II and possibly anglerfish peptide Y. Using [125I]D-Tyr-Val-Gly as substrate, a peptidyl-glycine alpha-amidating monooxygenase (PAM) was detected in islet secretory granule lysates. The enzyme is active between pH 6.0 and 8.5 and exhibits maximal activity near pH 7.0. The islet PAM requires Cu2+, ascorbate, and molecular oxygen for activity. Other divalent metal ions and redox cofactors were tested and found to be inactive in the assay. Even though added Cu2+ and ascorbate are required for detecting islet PAM activity, when these factors were incubated with substrate in the absence of secretory granule lysate, no activity was observed. It was also found that the addition of higher than optimal concentrations of either Cu2+ or ascorbate inhibited amidating activity. The results demonstrate that a PAM is present in secretory granules of anglerfish islet tissue. The characteristics of the islet PAM are similar to those of PAMs identified and characterized in other tissues which produce bioactive C-terminally amidated peptides.
authors.forename: R B
authors.initials: RB
authors.lastname: Mackin
authors.forename: J M
authors.initials: JM
authors.lastname: Flacker
authors.forename: J A
authors.initials: JA
authors.lastname: Mackin
authors.forename: B D
authors.initials: BD
authors.lastname: Noe
mesh_terms: D000818:Animals; D001205:Ascorbic Acid / Q000502:physiology; D003300:Copper / Q000502:physiology; D003594:Cytoplasmic Granules / Q000201:enzymology*; D004789:Enzyme Activation; D005399:Fishes / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D007515:Islets of Langerhans / Q000201:enzymology*; D006899:Mixed Function Oxygenases; D009097:Multienzyme Complexes; D010084:Oxidation-Reduction; D010100:Oxygen / Q000502:physiology; D010105:Oxygenases / Q000378:metabolism*
chemical_list: D009097:Multienzyme Complexes; D003300:Copper; D006899:Mixed Function Oxygenases; D010105:Oxygenases; C035189:peptidylglycine monooxygenase; D001205:Ascorbic Acid; D010100:Oxygen