Biosynthesis of peptide neurotransmitters: studies on the formation of peptide amides.
A high proportion of peptide transmitters and peptide hormones terminate their peptide chain in a C-terminal amide group which is essential for their biological activity. The specificity of an enzyme that catalyses the formation of the amide was investigated with the aid of synthetic peptide substrates. With peptides containing l-amino acids the enzyme exhibited an essential requirement for glycine in the C-terminal position; amidation did not take place with peptides that had leucine, alanine, glutamic acid, lysine or N-methylglycine at the C-terminus and a peptide extended by the attachment of lysine to the C-terminal glycine did not act as a substrate. Amidation did occur with a peptide containing C-terminal D-alanine but no reaction was detected with peptides having C-terminal, D-serine or D-leucine. In tripeptides with a neutral amino acid in the penultimate position, amidation, took place readily but the reaction was slower when this position was occupied by an acidic or a basic residue. A series of overlapping peptides with C-terminal glycine, based on partial sequences of calcitonin, underwent amidation at similar rates, indicating that the amidating enzyme recognizes only a limited sequence at the C-terminus of its substrates. The results provide evidence that the amidating enzyme has a highly compact substrate binding site.
Bradbury
A F
AF
National Institute for Medical Research, London, U.K.
Smyth
D G
DG
D000595:Amino Acid Sequence; D000818:Animals; D002250:Carbon Radioisotopes; D005998:Glycine; D007202:Indicators and Reagents; D007700:Kinetics; D008297:Male; D009479:Neuropeptides / Q000096:biosynthesis*; D018377:Neurotransmitter Agents / Q000096:biosynthesis*; D009842:Oligopeptides / Q000138:chemical synthesis; D010902:Pituitary Gland / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D013379:Substrate Specificity; D013552:Swine; D013961:Thyroid Gland / Q000201:enzymology*
D002250:Carbon Radioisotopes; D007202:Indicators and Reagents; D009479:Neuropeptides; D018377:Neurotransmitter Agents; D009842:Oligopeptides; D005998:Glycine