Peptidylglycine alpha-amidating enzyme (alpha-AE) can be used in an in vitro reaction to convert C-terminal glycine-extended peptides to peptide hormones with a C-terminal amino acid amide. Structure-activity data for 45 bioactive peptides show that the C-terminal amide is required for the full biological activity of most amidated peptide hormones. These data emphasize the role alpha-AE can have in amidated peptide production.