cDNA cloning and antibacterial activities of cecropin D-like peptides from Agrius convolvuli.
We have characterized full-length cDNAs encoding two isoforms of agriusin, cecropin D-like antibacterial peptide, present in the hemolymph of the immunized Agrius convolvuli larvae. The cloned cDNAs of agriusins 1 and 2 contain 331 and 329 bp, respectively. The nucleotide sequencing of cDNAs showed that they encode 62 amino acids, whose mature portion was deduced to consist of 38 amino acid residues with over 94% sequence identity. In the sequence homology search, mature agriusin 1 showed over 86 and 71% amino acid sequence homology with bactericidin 4 from Manduca sexta and cecropin D from Hyalophora cecropia, respectively. Since it was demonstrated from the deduced amino acid sequences that the C-terminal residues of agriusins are followed by a Gly residue, two types of synthetic agriusin 1 (syn-agriusin 1 amide and acid) were prepared to verify if natural agriusin 1 is C-terminally amidated. From acid-urea PAGE and reversed phase HPLC profiles to compare two synthetic peptides, we could confirm that the C-terminal amino acid residue of natural agriusin 1, like several cecropins so far identified, is amidated. Finally, our antibacterial assay performed with two syn-agriusins 1 revealed that there is little difference between antibacterial activities of both peptides against Gram-positive and Gram-negative bacteria.
Department of Life Science, Hoseo University, Asan City, Choongnam-Do, South Korea.
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D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000302:isolation & purification / Q000494:pharmacology*; D001412:Bacillus subtilis / Q000187:drug effects; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D019476:Insect Proteins / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D016982:Micrococcus luteus / Q000187:drug effects; D008969:Molecular Sequence Data; D009036:Moths; D010455:Peptides / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D020033:Protein Isoforms; D012486:Salmonella typhimurium / Q000187:drug effects
D000900:Anti-Bacterial Agents; D018076:DNA, Complementary; D019476:Insect Proteins; D010455:Peptides; D020033:Protein Isoforms; C428305:agriusin 1 protein, Agrius convolvuli; C428306:agriusin 2 protein, Agrius convolvuli; C036149:cecropin D protein, Hyalophora cecropia