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Rapid fluorimetric assay for the detection of the peptidyl alpha-amidating enzyme intermediate using high-performance
A P Consalvo1, S D Young, D J Merkler
1Department of Analytical Protein and Organic Chemistry, Unigene Laboratories, Inc., Fairfield, NJ 07004.
Journal of Chromatography
|August 21, 1992
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.