Molecular cloning of a peptidylglycine alpha-hydroxylating monooxygenase from sea anemones.
Cnidarians are the lowest animal group having a nervous system. The primitive nervous systems of cnidarians produce large amounts of a variety of neuropeptides, of which many or perhaps all are amidated at their C terminus. In vertebrates, peptide amidation is catalyzed by two enzymes acting sequentially, peptidyl-glycine alpha-hydroxylating monooxygenase (PHM) and peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL). In mammals both enzymatic activities are contained within a bifunctional protein that is coded for by a single gene. Using PCR and degenerated oligonucleotides derived from conserved regions of PHM, we have now cloned a PHM from the sea anemone Calliactis parasitica showing 42% amino acid sequence identity with rat PHM. Among the conserved (identical) amino acid residues are five histidine and one methionine residue, which bind two Cu2+ atoms that are essential for PHM activity. No cDNA coding for PAL could be identified, suggesting that sea anemone PAL is coded for by a gene that is different from the sea anemone PHM gene, a situation similar to the one found in insects. This is the first report on the molecular cloning of a cnidarian PHM.
Hauser
F
F
Department of Cell Biology and Anatomy, University of Copenhagen, Denmark.
Williamson
M
M
Grimmelikhuijzen
C J
CJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D005998:Glycine / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000235:genetics*; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics*; D009420:Nervous System / Q000201:enzymology; D009479:Neuropeptides / Q000096:biosynthesis; D010105:Oxygenases / Q000235:genetics*; D010455:Peptides / Q000378:metabolism; D012615:Sea Anemones / Q000201:enzymology / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D009097:Multienzyme Complexes; D009479:Neuropeptides; D010455:Peptides; D006899:Mixed Function Oxygenases; D010105:Oxygenases; C035189:peptidylglycine monooxygenase; D005998:Glycine