Characterization and cDNA cloning of a cecropin-like antimicrobial peptide, papiliocin, from the swallowtail butterfly, Papilio xuthus.
Cecropin is a well-studied antimicrobial peptide that is synthesized in fat body cells and hemocytes of insects in response to hypodermic injury or bacterial infection. A 503 bp cDNA encoding for a cecropin-like peptide was isolated by employing annealing control primer (ACP)-based differential display PCR and 5'-RACE with immunized Papilio xuthus larvae. The open reading frame of the isolated cDNA encoded for a 62-amino acid prepropeptide with a putative 22-residue signal peptide, a 2-residue propeptide, and a 38-residue mature peptide with a theoretical mass of 4060.89 Da. The deduced amino acid sequence of the peptide evidenced a significant degree of identity with other lepidopteran cecropins. This peptide was named papiliocin. RTPCR results revealed that the papiliocin transcript was detected at significant levels after injection with bacterial lipopolysaccharide (LPS). On the basis of the deduced amino acid sequence of papiliocin, a 38-mer mature peptide was chemically synthesized via the Fmoc method, and its antimicrobial activity was analyzed. The synthetic papiliocin peptide evidenced a broad spectrum of activity against fungi, Gram-positive and Gram-negative bacteria, and also evidenced no hemolytic activity against human red blood cells.
Kim
Seong Ryul
SR
Department of Agricultural Biology, National Academy of Agricultural Science, Rural Development Administration, Suwon, 441-100, Korea.
Hong
Mee Yeon
MY
Park
Seung Won
SW
Choi
Kwang Ho
KH
Yun
Eun Young
EY
Goo
Tae Won
TW
Kang
Seok Woo
SW
Suh
Hwa Jin
HJ
Kim
Iksoo
I
Hwang
Jae Sam
JS
D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D001483:Base Sequence; D002080:Butterflies / Q000235:genetics* / Q000378:metabolism; D002176:Candida albicans / Q000187:drug effects / Q000254:growth & development; D054807:Cecropins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D020869:Gene Expression Profiling; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000254:growth & development; D006094:Gram-Positive Bacteria / Q000187:drug effects / Q000254:growth & development; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D023181:Antimicrobial Cationic Peptides; D054807:Cecropins; D018076:DNA, Complementary; D019476:Insect Proteins; C551727:papiliocin protein, Papilio xuthus