title: Rapid fluorimetric assay for the detection of the peptidyl alpha-amidating enzyme intermediate using high-performance liquid chromatography.
abstract: Peptidylglycine alpha-amidating enzyme catalyzes the conversion of glycine-extended peptides to their corresponding amidated peptides via a stable alpha-hydroxyglycine intermediate. Using a new rapid fluorimetric reversed-phase high-performance liquid chromatographic assay, we have demonstrated that the substrate and product of the amidation reaction, as well as both stereoisomers of the alpha-hydroxyglycine intermediate, can be separated and detected in quantities as low as 1 pmol. The method is highly reproducible and requires less than 11 min for separation and quantification.
authors.affiliation: Department of Analytical Protein and Organic Chemistry, Unigene Laboratories, Inc., Fairfield, NJ 07004.
authors.forename: A P
authors.initials: AP
authors.lastname: Consalvo
authors.forename: S D
authors.initials: SD
authors.lastname: Young
authors.forename: D J
authors.initials: DJ
authors.lastname: Merkler
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D003619:Dansyl Compounds; D006899:Mixed Function Oxygenases / Q000032:analysis*; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D013050:Spectrometry, Fluorescence
chemical_list: D003619:Dansyl Compounds; D009097:Multienzyme Complexes; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase