Identification of a novel type of processing sites in the precursor for the sea anemone neuropeptide Antho-RFamide (<Glu-Gly-Arg-Phe-NH2) from Anthopleura elegantissima.
Neuropeptides are synthesized as large precursor proteins that undergo posttranslational cleavages and modifications to produce bioactive peptides. Here, we have cloned two closely related precursor proteins for the sea anemone neuropeptide Antho-RFamide (<Glu-Gly-Arg-Phe-NH2) from Anthopleura elegantissima. The first precursor (435 amino acids long) contains 13 copies of immature Antho-RFamide (Gln-Gly-Arg-Phe-Gly) and nine other, Antho-RFamide-related neuropeptide sequences that are in the C-terminal part of the protein. The second precursor (429 amino acid residues) harbors 14 copies of immature Antho-RFamide and eight other related peptide sequences. Each copy of Antho-RFamide or Antho-RFamide-related peptide is followed, at its C-terminal side, by a single Arg residue, which is an established signal for posttranslational cleavage. At the N terminus of each Antho-RFamide sequence, however, basic residues are lacking, and instead one or more acidic residues occur. These acidic residues are the cleavage sites for a new type of processing enzyme occurring in neurons. This enzyme could either be an amino- or endopeptidase hydrolyzing at the C-terminal side of Asp or Glu residues. The N-terminal regions of the two precursor proteins harbor eight copies of the putative neuropeptide sequence Pro-Gln-Phe-Trp-Lys-Gly-Arg-Phe-Ser and three additional, closely related sequences. The total number of all established and putative neuropeptides that may be cleaved from the precursors is 33. Thus, the Antho-RFamide precursors beong to the most complex peptide precursor proteins known so far.
Schmutzler
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Center for Molecular Neurobiology (ZMNH), University of Hamburg, Federal Republic of Germany.
Darmer
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Diekhoff
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Grimmelikhuijzen
C J
CJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D005796:Genes; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives; D012333:RNA, Messenger / Q000235:genetics; D012615:Sea Anemones / Q000737:chemistry*; D016415:Sequence Alignment
D009479:Neuropeptides; D012333:RNA, Messenger; C060288:4-pyroglutamyl-glycyl-arginyl-phenylalaninamide; D004247:DNA; D011761:Pyrrolidonecarboxylic Acid