Multipeptide precursor structure of acaloleptin A isoforms, antibacterial peptides from the Udo longicorn beetle, Acalolepta luxuriosa.
We previously purified acaloleptin A1, A2, and A3, antibacterial peptides that are produced in the larval hemolymph of Acalolepta luxuriosa (Udo longicorn beetle). In this study, we performed cDNA cloning. The cDNA sequence showed a predicted acaloleptin A precursor that consisted of five acaloleptin A isoforms. Four (isoforms 1, 2, 3 and 4) of the five isoforms of the acaloleptin A precursor had high-level sequence identities with each other, but the N-terminal region of isoform 5 differed from those of the other acaloleptin A isoforms. Northern and Western blot analyses showed that acaloleptin A isoforms were mass-produced soon after bacterial inoculation. Finally, we purified isoform 5 from hemolymph of the immunized larvae. Isoform 5, unlike acaloleptin A1, A2 and A3, showed antimicrobial activities against a Gram-positive bacterium, Micrococcus luteus and a fungus, Magnaporthe grisea. These results suggest that the multipeptide structure of the acaloleptin A precursor allows A. luxuriosa high-level production of antibacterial peptides and resistance to a wide range of microorganisms.
Imamura
Morikazu
M
Department of Applied Biological Science, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Saiwai-cho, Fuchu, Tokyo 183-8509, Japan.
Wada
Sugino
S
Ueda
Kenjiro
K
Saito
Ayaka
A
Koizumi
Nobuo
N
Iwahana
Hidenori
H
Sato
Ryoichi
R
D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D001483:Base Sequence; D001798:Blood Proteins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D015152:Blotting, Northern; D015153:Blotting, Western; D003001:Cloning, Molecular; D001517:Coleoptera / Q000235:genetics* / Q000378:metabolism / Q000382:microbiology; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005216:Fat Body / Q000378:metabolism; D005658:Fungi / Q000187:drug effects / Q000254:growth & development; D020869:Gene Expression Profiling; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000254:growth & development; D006094:Gram-Positive Bacteria / Q000187:drug effects / Q000254:growth & development; D006458:Hemolymph / Q000378:metabolism; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D007814:Larva / Q000235:genetics; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D011498:Protein Precursors / Q000235:genetics*; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid
D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D018076:DNA, Complementary; D019476:Insect Proteins; D020033:Protein Isoforms; D011498:Protein Precursors; C118274:acaloleptin A protein, Acalolepta luxuriosa